Bactericidal silver surfactant delivery into coating and polymer compositions

ABSTRACT

Disclosed are surfactant compounds and compositions that are antimicrobial. Also provided are polymeric compositions incorporating the surfactant compounds. The polymeric compositions may be used to form antibacterial coatings on surfaces.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application Ser.No. 60/958,836, filed Jul. 9, 2007, which is hereby incorporated byreference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

This invention was made with U.S. Government support under Grant No.DAAE07-03-C-L127 awarded in part by the United States ArmyTank-automotive and Armaments Command (TACOM). The government hascertain rights in the invention.

TECHNICAL FIELD

The invention relates generally to surfactants, more particularly, toanionic surfactants, and to anionic polyelectrolytes. More particularly,this invention relates to anionic surfactants and anionicpolyelectrolytes that are monodentate ligands for cations such as silveror sodium ions. This invention further relates to anionic surfactantsthat form reverse micelles and reverse microemulsions.

INTRODUCTION

Antimicrobial compositions are used in the health care industry, foodservice industry, meat processing industry, and, of course, byindividual consumers. Such widespread use of these compositions isindicative of the importance placed on controlling bacteria and othermicroorganism populations.

In general, antimicrobial agents are directed at bacteria, viruses, andfungi. Most agents, however, generally have a limited spectrum ofactivity. For example, bactericidal agents typically are not fungicidal,while fungicidal agents typically are not bactericidal. Quaternarysurfactants and agents have known antimicrobial properties. However,many irritate human skin and are unsuitable for many antimicrobialformulations. Anionic surfactants, although useful for washing or soapformulations, are considered, with a few exceptions, to lackantimicrobial activity.

The need for new disinfectant compositions is increasing in view of thedevelopment of methicillin resistant Staphylococcus aureus (MRSA). Theneed for new disinfectant compositions is increasing in view of thedevelopment of benzalkonium chloride resistant organisms (NobuyoshiAkimitsu et al., Antimicrobial Agents and Chemotherapy, December 1999,p. 3042-3043).

Ritu Bansal-Mutalik and Vilas G. Gaikar reported effects of reversemicelles on extractions from Escherichia coli ((2003) Enzyme andMicrobial Technology 32: 14-26). Water-in-hexane macro- andmicro-emulsions stabilized by sodium bis-(2-ethylhexyl)sulfosuccinate(NaAOT) were used for selective permeabilization of E. coli cells toextract penicillin acylase (3.5.1.11). Various organic solvents andsurfactants were compared for the yield and purification of the enzyme.Recoveries up to 144% with respect to sonication and specific activityof the enzyme up to 26 units/mg have been achieved. Due to lowsolubilities of NaAOT and aliphatic hydrocarbons in water, the recoveredenzyme was free from contaminants. A possible mechanism for cellpermeabilization and enzyme purification by reverse micellar treatmentwas proposed.

SUMMARY

In accordance with the invention, provided are antimicrobialcompositions or coating compositions comprising an anionic surfactant oran anionic polyelectrolyte. The invention further provides antimicrobialpolymeric compositions.

In another embodiment, the invention provides a method of making anantibacterial polymer coating.

In yet another embodiment, the invention provides an article ofmanufacture comprising an anionic surfactant or an anionicpolyelectrolyte.

In a further embodiment, the invention provides a method of treating aninfection or burn comprising administration of a composition comprisingan anionic surfactant or an anionic polyelectrolyte.

Other aspects of the invention will become apparent by consideration ofthe detailed description and accompanying drawings.

DETAILED DESCRIPTION

In accordance with the invention, anionic surfactants and anionicpolyelectrolytes are provided for delivering cations such as silver ion(Ag⁺) into various compositions. The invention further providescompositions including anionic surfactants or anionic polyelectrolytesand silver ion compositions that have antimicrobial properties. It hassurprisingly been discovered that anionic surfactants and suitableanionic polyelectrolytes of the invention serve as monodentate ligandsfor ions such as Ag⁺ and facilitate the incorporation of the ions intononaqueous compositions to render the compositions antimicrobial,specifically, bactericidal.

In accordance with the invention, a class of anionic surfactants hasbeen discovered that is lethal to Gram positive bacteria.

For the following defined terms, these definitions shall be applied,unless a different definition is given in the claims or elsewhere inthis specification.

All numeric values herein are assumed to be modified by the term“about,” whether or not explicitly indicated. The term “about” generallyrefers to a range of numbers that one of skill in the art would considerequivalent to the recited value (i.e., having the same function orresult). In many instances, the term “about” may include numbers thatare rounded to the nearest significant figure.

Weight percent, percent by weight, % by weight, wt %, and the like aresynonyms that refer to the concentration of a substance as the weight ofthat substance divided by the weight of the composition and multipliedby 100.

As used in the specification and the appended claims, the singular forms“a,” “an,” and “the” include plural referents unless the content clearlydictates otherwise. Thus, for example, reference to a compositioncontaining “a compound” includes a mixture of two more compounds. Asused in this specification and the appended claims, the term “or” isgenerally employed in its sense including “and/or” unless the contentclearly dictates otherwise.

It also is understood that any numerical range recited herein includesall values from the lower value to the upper value. For example, if aconcentration range is stated as 1% to 50%, it is intended that valuessuch as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are expresslyenumerated in this specification. These are only examples of what isspecifically intended, and all possible combinations of numerical valuesbetween and including the lowest value and the highest value enumeratedare to be considered to be expressly stated in this application.

It is further understood that the invention is not limited in itsapplication to the details of preparation and arrangement of componentsset forth in the following description or illustrated in the followingexamples. The invention is capable of other embodiments and of beingpracticed or of being carried out in various ways. Also, it is to beunderstood that the phraseology and terminology used herein is for thepurpose of description and should not be regarded as limiting. The useof “including,” “comprising,” or “having” and variations thereof hereinis meant to encompass the items listed thereafter and equivalentsthereof as well as additional items. “Comprising” encompasses the terms“consisting of” and “consisting essentially of.” The use of “consistingessentially of” means that a composition or method may includeadditional ingredients or steps, but only if the additional ingredientsor steps do not materially alter the basic and novel characteristics ofthe claimed composition or method.

Anionic Surfactants and Anionic Polyelectrolytes

Compounds embodying the principles of the invention include anionicsurfactants and anionic polyelectrolytes. Such surfactants andpolyelectrolytes serve as monodentate ligands for silver ion and othermetal ions, such as sodium, and serve as vectors or sequestering agentsto carry silver ion into suitable solutions, often nonaqueous andhydrophobic solutions. A monodentate ligand is a ligand that forms onlyone bond with the central atom, which is usually a metal ion. The typeand structure chosen for the anionic surfactants and anionicpolyelectrolytes depends on the solvent chosen and the final molecularcomposition into which the silver ion is to be sequestered and in whichthe silver ion is to exhibit its antimicrobial activity. For example, ifthe final composition has a significant poly(ethylene oxide) content,surfactants such as alkyloligomericethyleoxidesulfates may beparticularly useful for incorporating silver ion. Many of the anionicsurfactants of the present invention are commercially available. Anionicsurfactants for particular applications may be easily prepared bychemical procedures well known in the art and by methods describedherein.

Anionic surfactants are amphiphilic ions that typically have ahydrophobic or solvophobic part (for example, a dodecyl hydrocarbon tailas a hydrophobic group) and a hydrophilic or solvophilic part (forexample, a sulfate group bearing a negative charge, and a counter ionsuch as any alkali metal ion, such as Na⁺, or some small ammonium ion orthe like). The surfactant is termed “anionic” because its anioniccomponent is amphiphilic and will tend to segregate to interfaces, suchas air/liquid, air/solid, and liquid/liquid interfaces. For example, atan oil/water interface, the dodecyl group would mostly solubilize on theoil side of the interface, and the sulfate group would locate on thewater side of the interface, along with the counter cation.

The syntheses of surfactants such as soaps, ether carboxylic acids,alkylarylsulfonates, alkane sulfonates, olefin sulfonates, alcoholsulfates, alcohol ether sulfates, sulfated glycerides, sulfated alkanolamides, isethionates, taurates, sarcosinates, N-acyl amino acids, andα-sulfo fatty acid methyl esters are well known in the art. Thesyntheses of most of these surfactants are described in Reactions andSynthesis in Surfactant Systems edited by John Texter (Marcel Dekker,New York, 2001) in Chapter 1 (Ansgar Behler et al., pp. 1-44).

Specially, soaps are obtained by saponification of triglycerides in thepresence of the desired alkali hydroxide (e.g., NaOH), by saponificationof fatty acid obtained from fats and oils, and by saponification offatty acid methyl esters derived from fats and oils.

Ether carboxylic acids are obtained from fatty alcohol ethoxylates byreaction with chloroacetic acid in the presence of NaOH, by terminaloxidation of the fatty acid ethoxylate over Pt/C and neutralization withNaOH, and by the addition of a vinylic system, such as acrylonitrile,which hydrolyzes in aqueous HCl to the carboxylic acid, which is thenneutralized with the desired metal hydroxide.

Alkylarylsulfonates are obtained by reacting an alkylaryl, such as alinear or branched alkylbenzene, with oleum, sulfuric acid, or gaseoussulfur trioxide, and after aging are neutralized with NaOH or otheralkali hydroxide.

Alkane sulfonates are obtained by contacting an alkane-water mixturewith sulfur dioxide gas and oxygen at 30-40° C. under UV irradiation bysulfoxidation to produce the alkyl radical (R.), that then producesRSO₂., and after coupling with oxygen RSO₂OO., and “chain transfer” withthe starting material, RH, RSO₂OOH, with the generation of R. to keepthe synthesis going. The RSO₂OOH then reacts with water and sulfurdioxide to give RSO₃H and sulfuric acid.

Olefin sulfonates, especially α-olefin sulfonates (AOS), are obtained byreacting the alkene with SO₃ to produce the sultone. The sultoneabstracts a hydrogen atom from the carbon adjacent to the sultone (tothe original double bond) carbon to which the oxygen added to yield thealkenyl sulfonate. Hydrolysis converts any minor sultones of larger ringsize to the corresponding γ-hydroxy (and higher) alkane sulfonic acids.Neutralization with the desired hydroxide (e.g., NaOH) yields the olefinsulfonate.

α-Sulfo fatty acid methyl esters are obtained from fatty acid methylesters reacted with a 20% excess of SO₃ in air. The SO₃ initiallyinserts into the ester linkage but ends up inserted on the alpha carbonto produce the sulfonic acid. Neutralization with NaOH produces theα-sulfo fatty acid methyl ester.

Alcohol sulfates and alcohol ether sulfates are obtained by reacting thealcohol (X—OH) with 2 equivalents of sulfur trioxide gas to obtain thepryosulfate, X—OSO₂OSO₃H. The pyrosulfate then reacts with anotheralcohol, X—OH, to give two equivalents of the desired sulfate.Neutralization with alkali produces the desired salt.

Sulfated glycerides are obtained by reacting glycerol with oleum toobtain glycerol trifulfuric acid half-ester. Two moles of thishalf-ester are reacted with one of a triglyceride, and through a type oftransesterification one obtains 3 moles of monoglyceride sulfuric acidhalf-ester and three moles of sulfuric acid. Neutralization with alkaliyields the sulfate glyceride.

Sulfated alkanol amides are obtained by reacting the alcohol amide(X—OH) with 2 equivalents of sulfur trioxide gas to obtain thepyrosulfate, X—OSO₂OSO₃H. The pyrosulfate then reacts with anotheralcohol amide, X—OH, to give two equivalents of the desired sulfate.Neutralization with alkali produces the desired salt. Amide ethersulfates are obtained similarly if the amide ethoxylate is available.Otherwise an alkanol amide is ethoxylated and then reacted with sulfurtrioxide.

Isethionates are obtained by condensation of a fatty acid or othercarboxylate with sodium isethionate in the presence of an esterificationcatalyst at 200° C. or above. Alternatively, an acid chloride is reactedwith sodium isethionate to good yield after the HCl by product isremoved.

Taurates are similarly condensed with fatty acid chlorides or othercarboxylate chlorides to yield the taurate and the HCl by product. Thecarboxylic forms may be reacted directly at high temperature underesterification (amidation) conditions with water as the byproduct fromcondensation with the secondary amine.

Sarcosinates are most readily obtained by condensing the sodium form ofsarcosinic acid with fatty acid chlorides or with other acid chlorides,followed by removal of the HCl produced.

N-acyl amino acids can similarly be obtained by condensing the desiredacid chloride with the sodium form of the amino acid after removal ofthe HCl.

Phosphoric acid monoesters are obtained by reacting polyphosphoric acidwith 3 equivalents of alcohol, ROH, to obtain 3 equivalents of themonoester and 3 equivalents of phosphoric acid. Phosphorous pentoxide(P₄O₁₀) when reacted with 3 equivalents of water produces polyphosphoricacid, H₆P₄O₁₃. The monoesters are also obtained in 33% yield whenphosphorous pentaoxide is reacted with 6 moles of ROH. Here 4 moles ofdiester are obtained as the predominant product.

Polyelectrolytes that incorporate monodentate ligands for ions such assilver ions include carboxylates, sulfates, sulfonates, and phosphatesand are useful anionic polyelectrolytes of the present invention.Polystyrenesulfonate and copolymers of styrene sulfonate may suitablyincorporate silver ion into solutions and compositions of the presentinvention. Selection of co-monomers can be made so as to make the silverexchanged copolymer compatible with the final polymeric or coatingcomposition. Poly(sodium 4-styrenesulfonate) (PSS, M_(w)=70,000 g mol⁻¹)is available from Sigma-Aldrich (St. Louis, Mo., U.S.A). Similarlypolyvinylsulfate and copolymers of vinylsulfate are anionicpolyelectrolytes of the present invention. Poly(potassium vinyl sulfate)(PVS, (C₂H₃KO₄S) 162.1)_(n), n>1500), can be obtained from Wako PureChemical Industries, Ltd. (Osaka, Japan).

Varying co-monomer to achieve desired comparability and performanceproperties is well understood in the art. A statistical copolymer(Szczepan Zapotoczny, Monika Golonka, and Maria Nowakowska. Langmuir2008, 24, 5868-5876) of sodium p-styrenesulfonate (SSS) and2-vinylnaphthalene (VN) of molecular weight 150,000 g/mol is synthesizedusing the free-radical polymerization of the appropriate mixture of themonomers in degassed dimethyl sulfoxide (DMSO) solution for 21 h at 60°C. using 2,2′-azobis(2-methylpropionitrile) (AIBN, 0.1 mol %) as aninitiator. The content of VN monomer in the reaction mixtures is set to50 mol %, and the composition of the resulting copolymer is found using¹H NMR spectroscopy and elemental analysis to be almost the same (48 mol%) as the feed ratio. The resulting copolymer is purified by dialysis(Fisher, cellulose tubing, cutoff 12,000-14,000 g/mol; from FisherScientific, Waltham, Mass., U.S.A.) and subsequently freeze-dried.

Additionally polyacrylate (PA) and polymethacrylate are excellenthomopolymers for complexing with ions such as silver ion and forcarrying them into various substrates and compositions of the presentinvention. The natural product, i-type Carrageenan (CAG), is availablefrom Sigma-Aldrich (St. Louis, Mo., U.S.A.) as the sodium salt.

Water insoluble sulfonated poly(sulfone) sodium salts (SPSF) withdifferent ionic exchange capacities (IEC) and weight average molecularweights (MW) (IEC/MW) 0.85/94,000 g/mol and 0.65/83,000 g/mol) may besulfonated by an SO₃-TEP complex (2:1) by a method according to Byun etal. (J. Appl. Polym. Sci. 2000, 76, 787.).

Dextran sodium sulfate (DxS) is available commercially fromSigma-Aldrich (St. Louis, Mo., U.S.A.).

Structures (Morgan et al., Langmuir 2007, 23, 230-240) of anionicpolymers (SA) poly(sodium 2-acrylamido-2-methylpropane sulfonate)P(AMPS), (WA) poly(sodium 3-acrylamido-3-methylbutonate) P(AMBA), (B)poly(sodium3-acrylamido-3-methylbutonate-b-sodium-2-acrylamido-2-methylpropanesulfonate) P(AMBA-b-AMPS), (R) poly-(sodium3-acrylamido-3-methylbutonate-r-sodium-2-acrylamido-2-methylpropanesulfonate)P(AMBA-r-AMPS), homo and block copolymers of AMPS and AMBA are preparedusing a method similar to that reported (Sumerlin, B. S.; Lowe, A. B.;Thomas, D. B.; McCormick, C. L. Macromolecules 2003, 36, 5982-5987.).Briefly, homopolymers of AMPS and AMBA were prepared in water at 70° C.with V-501 as the initiator and CTP as the RAFT chain-transfer agent.The [CTA]/[V-501] ratio may be a 5:1 mole basis, with [CTA]) 2.54 ×10⁻⁴mol and [V-501]) 5.07×10⁻⁵ mol. To ensure that the acid functional groupon the monomer is neutralized, the pH of the polymerization solution maybe adjusted to about 8.4 by the addition of NaOH. Themonomer/chain-transfer agent, [M]/[CTA], ratios are chosen such that, ata quantitative conversion, number average degrees of polymerization(DPn) of 73 and 93 are attained, with [M] at 0.022 and 0.026 mol,respectively. Polymerizations may be conducted under nitrogen in a 22 mLreaction vial equipped with a magnetic stir bar. Polymers are purifiedby dialysis against deionized water and isolated by lyophilization.

Reverse-Micelle Forming Anionic Surfactants

Compounds embodying the principles of the invention include anionicsurfactants that form reverse micelles. Such anionic surfactants may bedouble-tailed or otherwise favor organic solvent solubilization overaqueous solubilization, and include sulfosuccinate diesters,sulfosuccinate monoesters, sulfosuccinimates, sulfo gluconate diesters,phosphoric acid monoesters, phosphoric acid diesters, and anionic Geminisurfactants. If an article in which the surfactant is to be incorporatedis a poly(2-ethylhexylmethacrylate) polymer or copolymer, sodiumbis(2-ethylhexyl)sulfosuccinate is a good choice of anionic surfactantbecause the surfactant is soluble in the liquid monomer solution and isreasonably soluble in the resulting polymer.

Sulfosuccinate diesters are most easily derived from maleic acidanhydride by reacting with an excess of the desired alcohol at 50-100°C. in the presence of a solvent (tolune, xylenes) to azeotropicallyremove the water produced and a catalyst, such as p-toluenesulfonicacid, followed by sulfonation in an aqueous solution of sodium hydrogensulfite. Drying produces the solid. Sulfosuccinate monoesters aresimilarly obtained from the desired alcohol, but without solvent at70-100° C. using a 1:1 stoichiomery.

Sulfosuccinamate diesters are derived from maleic acid anhydride byreacting with the desired amine in the presence of a catalyst, such asp-toluenesulfonic acid, followed by sulfonation in an aqueous solutionof sodium hydrogen sulfite. Drying produces the solid. Sulfosuccinamatemonoesters are similarly obtained from the desired amine, but using a1:1 stoichiomery.

The preparation of ammonium and tetrapropylammonium salts ofbis(2-ethylhexyl)sulfosuccinate have been reported by Eastoe et al.(Langmuir 1993, 9, 2820-2824). A cation exchange resin (Amberlite IR 120Plus, 2.0 mequiv g) is converted to its H⁺ form by equilibration with1.0 M HCl. A 50 mL sample of a 1.0 M ethanolic solution of Na(AOT) ispassed through the column and the H⁺ form of the surfactant produced.The first 20 mL of eluant is discarded; the remainder is reacted in situto pH 7.0 with an aqueous solution of the quaternary ammonium hydroxide(Sigma-Aldrich, St. Louis, Mo., U.S.A.). During the reaction the pH ismaintained in the range 5.0-8.5 and the solution continually stirred.The surfactant is obtained by evaporating the organic phase to dryness(Buchi rotary evaporator) at 35° C.; residual water is removed in avacuum oven at 35° C. for 3 days. The dry products are obtained as whitewaxy solids.

The straight chain and branched compounds may be prepared in a similarway from the appropriate starting alcohol, as described in Sandrine Naveand Julian Eastoe, and Jeff Penfold, Langmuir 2000, 16, 8733-8740).Butan-1-ol, pentan-1-ol, hexan-1-ol, heptanol-1-ol, octan-1-ol,2-ethyl-1-hexanol, 2-propyl-1-pentanol, 2,4,4-trimethyl-1-pentanol,3,5,5-trimethyl-1-hexanol, 4-methyl-3-heptanol, and 6-methyl-2-heptanolmay be purchased from Sigma-Aldrich (St. Louis, Mo., U.S.A.). As anexample the synthesis of di-C₄SS is outlined below. Briefly, fumarylchloride (Avocado, 95%) is added dropwise to a stirred solution ofbutan-1-ol in dry tetrahydrofuran (THF; Aldrich, 99+% anhydrous). Thereaction mixture is then refluxed until TLC indicated completion. Afterrotary evaporation of THF, the mixture is dissolved in diethyl ether andwashed several times sequentially with 10% HCl and saturated NaHCO₃solutions. The ethereal extracts are dried over anhydrous MgSO₄ andfiltered, and diethyl ether is removed by rotary evaporation. Vacuumdistillation yields the pure diester, which is a yellow oil. This isthen dissolved in a 1:1 mixture of ethanol/water and refluxed with aslight excess of sodium metabisulfite and sodium sulfite (Avocado, 97%)to form the dibutyl sulfosuccinate. Soxhlet extraction with ethylacetate, followed by several centrifugation cycles in methanol, may beemployed to remove residual sodium salts. All the di-CnSS surfactantsmay be further recrystallized from methanol. The final products may bedried in a vacuum oven (50° C., 5 mbar) for at least 24 h, and thenstored in a desiccating cabinet over phosphorus pentoxide, in sealedvials.

Dickson et al. (Ind. Eng. Chem. Res. 2005, 44, 1370-1380) reported thesynthesis of the phosphate diester, di(2,4,4-TMC5)phosphate⁻NH₄ ⁺. Aflask containing 2,4,4-trimethylpentanol (6.11 mL) and triethylamine(6.0 mL) dissolved in 30 mL of diethyl ether is degassed with argon andcooled to 0° C. This solution is slowly cannulated into anargon-degassed solution at 0° C. containing 1.79 g of phosphorusoxychloride in 30 mL of diethyl ether. A white precipitate forms uponaddition. This solution is stirred overnight while slowly being warmedto room temperature. The triethylamine hydrochloride salt is filtered,washed with 50 mL of diethyl ether, and discarded. The diethyl ether andexcess triethylamine are removed via rotary evaporation, leaving ayellow oil. This oil is dissolved in 40 mL of acetonitrile followed byslow addition of 3 mL of distilled water with stirring. Afterapproximately 7 h, the excess water and acetonitrile are removed viarotary evaporation, providing 4.9 g of crude product. The oil isdissolved in chloroform and purified by being passed through a silicagel column, starting with chloroform as the mobile phase and changing toa 9:1 chloroform/methanol mixture. To produce 2,4,4-TMC5—PO₄ ⁻NH₄ ⁺, theresulting yellow oil is dissolved in 50 mL of ethanol, and a solution ofammonium hydroxide (0.65 mL of glacial NH₄OH in 10 mL of methanol) isadded dropwise to the solution and stirred overnight. A whitish solid,trichained phosphates, is filtered and discarded, and the filtrate wasconcentrated via rotary evaporation. The resulting solid is dissolved in100 mL methanol with 1 g of decolorizing carbon and stirred for 1 h. Thecarbon is filtered, and the solvent is removed via rotary evaporation.The crude product is dissolved in a minimum amount of methanol, andacetone precipitated the white gel-solid. To produce 2,4,4-TMC5—PO₄⁻N(CH₃)₄ ⁺, the yellow oil is dissolved in 50 mL of ethanol, and 6.76 mLof tetramethylammonium hydroxide (25 wt % in methanol) is added dropwiseand stirred overnight. No solids typically form. Consequently, thematerial is concentrated via rotary evaporation. The resulting gel-solidis dissolved in 100 mL of methanol with 1 g of decolorizing carbon andstirred for 1 h. The carbon is filtered, and the solvent is removed viarotary evaporation. The crude product is dissolved in a minimum amountof methanol, and acetone precipitated bright white crystals.

The synthesis of 1,2-dilauroyl-sn-glycero-3-phosphatidyluridine wasreported by Francesca Baldelli Bombelli, Debora Berti, Uwe Keiderling,and Piero Baglioni, (J. Phys. Chem. B 2002, 106, 11613-11621).1,2-Dilauroyl-sn-glycero-3-phosphocholine may be purchased from AvantiPolar Lipids (Alabaster, Ala., U.S.A.), and its purity may be checked byTLC. DLPU may be synthesized starting from the correspondingphosphatidylcholine in a two-phase system according to the methodproposed by Shuto and co-workers (Chem. Pharm. Bull. 1988, 35, 209-217)and obtained as an ammonium salt.

The effect of carbon number has been studied (A. K. Chattopadhyay, D. O.Shah, and L. Ghaichao, Langmuir 1992, 8, 21-30) and example syntheseswere demonstrated (A. K. Chattopadhyay, U.S. Pat. No. 4,919,179,incorporated herein by reference) with three different series of doubletailed surfactants, viz. poly(isobutylene)succinic anhydride esterifiedwith di- and triethanolamine esters of n-alkylsuccinic anhydride andpoly(isobutylene)succinic anhydride esterified with((n-alkyloxy)propyl)diethanolamine. The primary hydrocarbon chain of thesurfactants comprises polyisobutylene of approximately 34 backbonecarbon number and the secondary hydrocarbon chain comprises n-alkylgroups varying from C₈ to C₂₀. Poly(isobutylene)succinic anhydride ofaverage molecular weight 1050 (supplied by Paramins Exxon, Linden, N.J.,U.S.A.) may be further purified by chromatographic separation in orderto avoid contaminations from free oils or poly(isobutylene) components.((n-Alkyloxy)propyl)diethanolamines of Tomah Products (Milton, Wis.,U.S.A), n-alkylsuccinic anhydride of Humphrey Chemicals (North Haven,Conn., U.S.A.), and reagent grade di- and triethanolamines (BDH) areused without any further purification. Surfactants of Series A may besynthesized by reacting diethanolamine with poly(isobutylene)succinicanhydride (PIBSA) followed by reaction with n-alkylsuccinic anahydride.With R₂=tetradecyl, 29.6 g of tetradecylsuccinic anhydride is heated toa 60-65° C. Then 10.5 g of diethanol amine is added dropwise to theanhydride with constant stirring. Completion of reaction and formationof esters is noted by IR spectroscopy by the disappearance of theabsorbance peak at 1790 ν⁻¹ due to anhydrides and appearance of theester peak at 1740 ν⁻¹. Further reaction between PIBSA and the adduct iscarried out by using all of the above reaction products together with167 g of PIBSA. The surfactant is produced as a 50% w/w solution inparaffin oil; and conversion to alkali salts is done by titration withMOH, where M⁺ is an alkali cation. Surfactants of Series B aresynthesized by reacting triethanolamine with n-alkylsuccinic anahydridefollowed by reaction with poly(isobutylene)succinic anhydride. Forexample, 26.8 g of dodecylsuccinic anhydride is heated to a temperaturein the range of 60-65° C. Then 14.9 g of triethanolamine is addeddropwise to the anhydride with constant stirring. Completion of reactionand formation of esters is noted by the same way as described for SeriesA. Further reaction between PIBSA and the adduct may be carried out byusing all of the above reaction products together with 167 g of PIBSA.The surfactant is produced as a 50% w/w solution in paraffin oil;conversion to alkali salts is done by titration with MOH, where M⁺ is analkali cation. Surfactants of Series C are synthesized by reacting((n-alkyloxy)propyl)diethanolamine with poly(isobutylene)succinicanhydride at 80° C. Conversion to alkali salts is done by titration withMOH, where M⁺ is an alkali cation.

Gemini surfactants consist of two surfactant moieties, linked togetherby a spacer unit such as 2, 3, 4, or more methylene groups. Geminianionic surfactants based on EDTA (Laurent Wattebled and AndréLaschewsky, Colloid Polym Sci (2007) 285: 1387-1393) such as thedidecyldimethyl amido derivative pictured below are obtained by reactionof secondary amines with EDTA anhydride. N-methyldodecylamine (3.20 g,16 mmol) and EDTA anhydride (2.05 g, 8 mmol) suspended in CH₃OH (50 ml)are reacted for 22 h at 40-45° C. The anhydride particles progressivelydisappear as the reaction progresses. After cooling of the sample toroom temperature the remaining particles are filtered off. The reactionmixture is evaporated to give a yellowish oil. Acetone is then addeduntil a white solid precipitates. The precipitate is filtered off and isfurther purified by dissolution in CHCl₃ and precipitation in acetone toyield a white powder. Finally, the intermediate product is neutralizedwith sodium hydroxide (1 M aq, 2 equivalents), and the obtained solutionis freeze-dried to give gemini surfactant in quantitative yield acolorless, hygroscopic powder.

Other substituents may be incorporated by simply varying the startingamine RNH₂ or R₁R₂NH, where R, R₁, and R₂ are alkyl, alryl, alkylaryl,or otherwise chosen to make the surfactant compatible with particularchoices of solvent, polymer, and final composition. Straight chain andbranched carbon chains of 6-18 carbon atoms are preferred for theprimary amines, and a total of 4 to 10 carbon atoms for each of R₁ andR₂ combined are preferred for the secondary amines when high solubilityin nonaqueous environments is desired. The cation may be varied bychoosing an appropriate hydroxide solution, such as LiOH, KOH, ortetramethylammonium hydroxide, etc.

Other anionic Gemini surfactants are described by S. K. Hait and S. P.Moulik (Current Science, Vol. 82, No. 9, 10 May 2002, pp. 1101-1111):

The following Gemini phosphate esters were reported by Menger and Littau(J. Am. Chem. Soc, 1991, 113, 1451-1452). They may be synthesized byfirst reacting α,α′-dibromo-p-xylene with dianionic alkyl phosphatemonoesters.

Menger and Littau also reported the photosensitive styryl Geminiphosphate ester prepared by phosphorylating the spacer diol with POCl₃,and then reacting the intermediate with long chain alcohols.

The following Gemini sulfate and sulfonates have ethylenoxide spacers.

TABLE 1

Properties of Gemini sulfates (A) and sulfonates (B) in comparison withsodium laurylsulfate and sodium laurylsulfonate. Compound^(a) Y CMC/mMγ_(cmc)/N m⁻¹ C₂₀/mM A —OCH₂ CH₂O— 0.013 27.0 0.001 C₁₂H₂₅SO₄Na — 8.239.5 3.1 B —O— 0.033 28.0 0.008 B —OCH₂ CH₂O— 0.032 30.0 0.0065 B—O(CH₂CH₂O)₂— 0.060 36.0 0.01 C₁₂H₂₅SO₃Na — 9.8 39.0 4.4

The double chain dicarboxylate forms vesicles (Jaeger, D. J. and Brown,E. L. G., Langmuir, 1996, 12, 1976-1980.). The reaction of threo diolester 4 with α-keto ester 5 gives diastereomeric diester ketals 2. Thismixture is converted into diacid ketals 3. A 1:1 mixture of surfactants1a and 1b is generated in situ by the dispersal of 3 into a pH 9.2 or10.7 KHCO₃—K₂CO₃ buffer. The potassium surfactants are more soluble thanthe sodium surfactants.

The triple-chained di-carboxylate below (Sumida, Y. et al., Chem.Commun., 1998, 2385-2386) can sequester metal ions effectively.

Syntheses of fluorinated phosphate diesters (S. Jason et al., Langmuir2004, 20, 1065-1072) F/F and H/F,

are outlined below:

To synthesize diH₈—PO₄ ⁻Na⁺[di(CF₃(CF₂)₄CH₂CH₂)phosphate⁻X⁺] (Jasper L.Dickson et al., Ind. Eng. Chem. Res. 2005, 44, 1370-1380), phosphorusoxychloride (1.79 mL) may be first added via syringe to 30 mL ofanhydrous diethyl ether, under argon. The mixture is cooled to 0° C.,and a cold solution of 1-octanol (6.05 mL) and triethylamine (5.82 g) in25 mL of diethyl ether is slowly added via cannula, giving whiteprecipitate, triethylamine hydrochloride salts. The solution is allowedto warm to room temperature and stirred under argon overnight. Thetriethylamine hydrochloride salts are filtered and washed with 50 mL ofdiethyl ether. The solvent and excess triethylamine are removed viarotary evaporation, and the product is dissolved in 40 mL ofacetonitrile and 1 mL of water and stirred overnight. The product iscollected via rotary evaporation and identified as the neutral phosphateusing NMR spectroscopy. After the product is dissolved in 40 mL ofethanol, a mixture of aqueous NaOH (0.17 ML, 50 wt %) dissolved in 25 mLof ethanol is added dropwise to the stirring phosphate solution. Afterbeing stirred overnight, the product is filtered, and acetone is addedto induce precipitation of a white salt, which is filtered and rinsedwith 30 mL of acetone.

Anionic surfactants according to the invention further include thefollowing, wherein M⁺ is a metal cation:

Silver Anionic Surfactants and silver Anionic Polyelectrolytes

Anionic surfactant salts according to the invention may be changed fromone cationic form to another cationic form, for example, from the sodiumto the silver form. Suitably, sodium surfactants may be converted toacid form using ion exchange or ion exchange chromatography, asdescribed in Petit et al. (1993) J. Phys. Chem. 97: 12974-12983, herebyincorporated by reference. Resins for the ion exchange may suitablyinclude protonated strong cationic exchange resins such as Lewatit SP1080 (LANXESS, Pittsburgh, Pa., U.S.A.). The acid form may then beloaded onto a weakly acidic macroporous cation exchange resin, such asBio-Rex 70 (BIO-RAD, Hercules, Calif., USA) loaded with silver ion. Aneluent of a 50/50 mixture of water and ethanol may be used. The finalproduct may be dried under vacuum. The efficiency of the conversion ofions may be determined by silver ion concentration titration usingVoPhard's method, with the precipitation of thiocyanate in the presenceof ammonium ferric sulfate as the indicator. Alternatively, a simplepotentiometric tirtration with standard chloride in methanol ormethanol/water may be done using a silver, silver halide, or silversulfide indicating electrode, and any suitable and convenient referenceelectrode, such as AgCl, with any suitable salt bridge.

An alternative approach to producing anionic surfactants in the silverform may be useful when the protonated or acid form of the surfactant isreadily available. The acid form, in any suitable solvent, is stirredwith an equivalent of silver oxide (silver hydroxide, silveroxyhydroxide). The by product is water.

Additionally, simple conversion from alkali metal form to the silver ionform may be accomplished by two phase ion exchange. The surfactant isdissolved in a convenient water immiscible solvent, such asdiethylether, and combined with an equal volume of 0.1 M aqueous AgNO₃in a separatory funnel. The alkali cation is extracted into the aqueousphase and the silver ion is extracted into the ether phase, sequesteredby the surfactant. This washing is repeated several times, and then theether phase is washed with water, and the phases are separated. Theeither phase is then reduced in volume on a rotary evaporator, and theremaining paste is dried in a vacuum oven at 25-50° C.

A similar method for effecting ion-exchange of cations in the anionicsurfactants and polyelectrolytes of the present invention is thefollowing method reported by Steytler et al. (Langmuir 1996, 12,1483-1489). Bis(2-ethylhexyl)phosphoric acid (HDEHP, 97%, Aldrich) isinitially purified according to the method of Partridge et al. (J.Inorg. Nucl. Chem. 1969, 31, 2587). The general method used to prepareall the multivalent di(2-ethylhexyl)phosphate (DEHP) metal saltsinvolved neutralization of HDEHP with the metal hydroxide. In the caseof divalent metals, M(OH)₂ is freshly prepared as a precipitate bymixing an aqueous solution of the metal nitrate (15 mL; 1 mol dm⁻³) withexcess aqueous sodium hydroxide solution (20 mL, 1 mol dm⁻³). The M(OH)₂precipitate is then filtered and thoroughly washed with water to removeexcess NaOH. The metal hydroxide is then added directly to a biphasicsystem comprising water (50 mL) and a solution of HDEHP in diethyl ether(25 mL, 1.1 mol dm⁻³) contained in a separating funnel. The resultingmixture is thoroughly shaken and left to phase separate overnight. Thelower aqueous phase is then removed, together with any excess M(OH)npresent at the interface, and the ether layer is repeatedly washed withfresh aliquots of water and reduced in volume to about 15 mL. TheM^(n+)(DEHP⁻)n salt is subsequently precipitated by the slow addition ofacetone to the ether solution with stirring. After completeprecipitation, the product is filtered and washed repeatedly withacetone. The M^(n+)(DEHP⁻)n salts are initially dried in a vacuum ovenat 40° C. for about 48 h and further dried by storing over P₂O₅ in avacuum desiccator until required.

Similar procedures applied to bis(2-ethylhexyl)sulfosuccinate aredescribed by Eastoe et al. (J. Phys. Chem. 1993, 97, 1459-1463). TheM⁺(AOT⁻) surfactants (M═K⁺, Rb⁺, and Cs⁺) may be prepared using themethod due to Hatton (E. B Leodidis,; T. A. Hatton, In Structure andReactivity in Reversed Micelles; M. P Pileni, Ed.; Elsevier: New York,1989; p 270.). Briefly, approximately equal volumes of a 1.0 mol dm⁻³aqueous solution of the metal nitrate (Sigman-Aldrich, St. Louis, Mo.,U.S.A.) with a 0.10 mol dm⁻³ heptane solution of Na(AOT) are shakentogether. On settling, a Winsor II microemulsion system may be formed:the organic upper phase is removed and shaken with fresh 1.0 mol dm⁻³metal nitrate twice more. The M⁺(AOT⁻) surfactant is then obtained byevaporating the organic phase to dryness (Buchi rotary evaporator). Thedry product is stored under vacuum over P₂O₅ (Aldrich) until used. Thesurfactants are analyzed for the extent of replacement of Na⁺ for Mn⁺using either UV/vis or atomic absorption (Varian Spectra AA-10)spectrophotometry. The amount of residual water (x) in the productM⁺(AOT⁻).xH₂O is assessed with a calibrated FTIR measurement of theintegrated area of the absorbance due to the O—H stretch (3700-3050cm⁻¹).

When the anionic polyelectrolyte is synthesized for a particularapplication, it may be expedient to use the acid monomers rather thanalkali or ammonium cation salts of the monomer. After synthesis iscomplete the acid polymer is mixed with an equivalent amount of silveroxide in a suitable solvent, where the conversion is quantitative. Thesolvent and water produced are then removed by vacuum drying.

Alternatively, starting with alkali metal salts of anionic copolymer ofstyrene sulfonate, sodium salt, and styrene (25 mol % styrenesulfonate), the copolymer is dissolved in toluene and ion exchanged intothe silver ion form by using the two-phase ion exchange processdescribed above. An equal volume of 0.1 M silver nitrate is added to thetoluene solution in a separatory flask, shaken, and exchanged twice morewith aqueous silver nitrate, then washed twice with deionized water. Thewater phase is separated, and the toluene solution is reduced in volume,and finally dried in a vacuum oven at 50° C. overnight.

Formation of the Composition

A particular advantage of the method of inclusion of the silver ionmonodentate ligand complexes of the present invention, in comparison toformulations of prior art silver nanoparticles and silver saltnanoparticles, is that the instant silver ion complexes dissolve in thesolutions of the present invention as a result of chemical free energyreductions. Inclusion of nanoparticulate dispersions of silver are atbest metastable, and extraordinary stabilization methods, nanoparticleformation methods, and high energy dispersing methods must be used tocreate useful formulations of such prior art silver sources. In thepresent invention, the silver surfactants and silver polyelectrolytessimply dissolve, as equilibration into the dissolved state is a negativefree energy (i.e., spontaneous) process.

The particular solvent or solvent mixture to use is selected in order tooptimize the method of application and the physical and chemical natureof the substrate to be contacted by the silver monodentate ligandcontaining solution. A prime consideration for applying silversurfactant and silver polyelectrolyte solutions of the invention is thatsaid solution will wet the substrate. In cases where the desired wettingis at first not obtained, an alternative surfactant resulting in lowersurface tensions of the solution may be used. Alternatively,fluorocarbon surfactants of the present invention may be added to thesolution, in silver ion complexed form or with any other suitablecation. Inclusion of such fluorocarbon containing surfactants in saidsolutions will be of particular help in obtaining satisfactory wettingon plastic and other polymeric surfaces and substrates, includingspecialized coatings and enamels that are normally difficult to wet.

Some useful solvents of the invention are listed in Table 2 along withtheir corresponding surface tensions at or near room temperature. Theselection of a solvent or solvent mixture may take account ofventilation requirements and possible safety hazards because of theflammability of some of the solvents. For example, water (sterile water)is a preferred solvent for delivering the surfactants of the inventionto mammalian tissues. Isopropanol and ethanol may be mixed with waterfor delivery to and coating on exterior essentially intact skin tissue.For applications to open wounds, burns, and surgical cavities, sterilewater and sterile aqueous saline are preferred in order to minimizeuntoward tissue dehydration.

TABLE 2 Useful Solvents and Their Surface Tensions. Solvent SurfaceTension (dyn/cm) Trifluoroacetic Acid 13.63 (24° C.) Pentane 15.48 (25°C.) Ethyl Ether 17.06 Hexane 17.91 (25° C.) Iso-Octane 18.77Acetonitrile 19.10 Methyl t-Butyl Ether  19.4 (24° C.) Heptane 20.30Triethylamine 20.66 Isopropyl Alcohol 21.79 (15° C.) Ethyl Alcohol 22.32Cyclopentane 22.42 Methanol 22.55 Isobutyl Alcohol 22.98 Acetone 23.32Methyl Isobutyl Ketone 23.64 n-Propyl Alcohol 23.70 n-Butyl Chloride23.75 Ethyl Acetate 23.75 Methyl Ethyl Ketone  24.0 (25° C.) n-ButylAlcohol 24.57 Cyclohexane 24.98 n-Butyl Acetate 25.09 Methyl n-PropylKetone 25.09 Tetrahydrofuran  26.4 (25° C.) o-Dichlorobenzene 26.84Chloroform 27.16 Dichloromethane 28.12 Toluene 28.53 o-Xylene 30.032-Methoxyethanol  31.8 (15° C.) Ethylene Dichloride 32.23 DimethylAcetamide 32.43 (30° C.) o-Dichlorobenzene 26.84 Chloroform 27.16Dichloromethane 28.12 Toluene 28.53 o-Xylene 30.03 2-Methoxyethanol 31.8 (15° C.) Ethylene Dichloride 32.23 Dimethyl Acetamide 32.43 (30°C.) Chlorobenzene 33.28 1,4-Dioxane 34.45 (15° C.) N,N-Dimethylformamide36.76 Pyridine 36.88 Propylene Carbonate 41.93 Dimethylsulfoxide 43  Water 72.8 

In some cases, commercial surfactant formulations may be required inorder to effect wetting of the solutions of the present invention onrelatively low surface energy substrates and surfaces. Table 3 lists aseries of hydrocarbon/fluorocarbon solvents, Vertel® solvents, availablefrom the DuPont Corporation through Micro Care Company (Bristol, Conn.,U.S.A.).

TABLE 3 Commercially Available FluoroHydrocarbon Solvents and TheirSurface Tensions. Surface Tension Solvent (dyn/cm) Vertel ® XF 14.1Vertel ® XM 14.1 Vertel ® XDA 14.1 Vertel ® XP10 14.1 Vertel ® XMS Plus14.9 MicroCare CF 15.0 Vertel ® XP 15.1 Vertel ® MCA 15.2 Vertel ® SMT15.5 Vertel ® MCA Plus 16.1 Vertel ® CCA 18.8 Vertel ® CMS 19.2 Vertel ®CHD 19.4

Methods of Applying the Composition

The solutions of the present invention are intended to be delivered ontoand into surfaces and substrates of materials so as to eradicate anybacteria already present on and in such surfaces and substrates by thebactericidal activity of the invention surfactants and complexes. It isalso anticipated that once applied to a surface or substrate thatprophylactic protection against bacteria coming to reside on or in saidsurfaces and substrates will continue for such time until coating isremoved.

Spraying the solutions of the invention onto and into surfaces andsubstrates may be done by any normal means of spraying. For example, inthe case of spraying into shoes, onto shirt underarm areas, and intosports uniforms between launderings, small (5-100 mL) hand held sprayersmay be used, whereby the aerosol is generated by finger pressure on anozzle/pump assembly. As the area and magnitude of area and materialsincreases, larger spray bottle may be used, for example wherein the pumpdriving the aerosol generation is pumped by a handle activated by fourof the fingers in a hand. If walls, ceilings, and their attachedfixtures are to be sprayed, larger volume pressurized sprayers arepreferred. Spraying is a suitable method for applying the solutions ofthe present invention to fabrics, particularly to moving webs of fabric.Arrays of spray nozzles can be used to coat the solutions onto and intothe fabric in a uniform manner.

Pouring is a suitable way to apply the solutions and fluids of thepresent invention to a small to large horizontal area. In the case ofsmall areas, the pouring method may be augmented with wiping, coating,or spinning. In the case of larger areas, the pouring method may beaugmented by squeegee blades of the type, for example, used in washingwindows. The contact edges of said squeegee blades may be modified withgrooves and slits so as to insure a given thickness of fluid remains onthe surface after passing the blade over the surface.

Where the volume or weight of solution or composition administered iscontrolled, metering may be used to apply the compositions of thepresent invention for inhalation treatments, where the amount of aerosoldosing the lungs may be important to control. Metering may also be usedto control flow and coating amounts in combination with pumping,filling, and coating methods of application.

Coating is a suitable application method for sheet materials and fabricssuitable for continuous web treatment. Doctor blade, x-hopper, slidehopper, gravure, and curtain coating are preferred methods of coatingthe solutions and fluid compositions of the present invention to sheetmaterials and fabrics. Dip coating is a suitable method for coatingsmall to intermediate sized object and materials. Dip coating is asuitable method for coating fabrics that are absorbent.

Pumping may be used to move the solutions and fluid compositions of thepresent invention from a reservoir or holding tank to a spray nozzle,x-hopper coating surface, slide hopper coating surface, or curtaincoating apparatus. Hand operated spraying may incorporate a finger orhand actuated pump.

Wiping using absorbent cloths, sponges, and other absorbent materialsthat infuse the solutions of the invention and then exude them onto acontacting surface when pressed against said surface may be used toapply the compositions of the invention to walls, counter tops,shelving, windows, window sills, tables, and other objects. On largersurfaces a volume may be applied with a sponge and then the appliedvolume is spread over a larger area by action of a hand operated blade.

Flowing is a suitable method of application when the invention solutionsare to be applied to a continuously moving web, such as in an operationof x-hopper or slide hopper coating are as in curtain coating. Anotherapproach may be where a large volume spray is directed along the top ofa wall area, and the rest of the wall is covered by allowing theimpacting solution to simply flow down the wall, wetting the lowerportions as the vertically flow proceeds.

Drying

After application of the solutions and fluids of the present invention,the surfaces and substrates to which these solutions and fluids havebeen applied may be dried. Evaporation is a suitable method of drying.In manufacturing environments where air quality and safety areparamount, temperature programmed drying, heating, and solvent recoverymay be important components to the overall process. Heating may benecessary when solvents with relatively high boiling points are used.

Surfaces and Substrates

The surfaces and substrates to which the solutions and compositions ofthe present invention range from those encountered in the practice ofmedicine with human tissue and in veterinary medicine with animaltissues of every type, including severely burned epidermis and dermistissues, open wounds in the category of scrapes and scratches, incisionsencountered in surgery, lung, nasal, and vaginal mucosal tissues,abdominal and thoracic tissues line the organs and cavities of theabdomen and chest cavity, bone fracture surfaces, and surfaces liningthe mouth, gums, ears, urethra, and skin surfaces, to the surfaces andsubstrates encountered in the interiors of homes, schools, hospitals,manufacturing plants, and other public places of business andgovernment.

Existing wound dressing materials of woven and sheet form includesurfaces and substrates to which the solutions and compositions of thepresent invention can be applied. The threads and strands used in makingsuch woven materials may be coated with the solutions and compositionsof the present invention by dip coating during the spinning and weavingprocesses during manufacture. Alternatively, the compounds of thepresent invention may be incorporated into the bulk of the syntheticmaterial prior to spinning such polymers into strands and threads to bewoven into would dressings.

All clothing articles that come in contact with sweat and bacterianormally populating human skin are objects benefiting from the presentinvention. Fabrics and shoes may be treated with the solutions andcompositions of the present invention by spraying, dip coating, andwiping. Alternatively, the materials comprising the contact surfaces ofclothing articles may incorporate the surfactants and polymers of thepresent invention during their manufacture, so that they exudebactericidal activity over a continuing time interval. Applications totextiles in general may be made either to the final product by one ormore application methods of the present invention or they may beincorporated directly into polymeric compositions used to preparesynthetic fabrics.

Surface and substrates in the home are also anticipated as targets forthe solutions and fluid compositions of the present invention. Wall andceiling, as well as all objects and fixtures attached thereto, arepossible places for bacteria and microbes to inhabit. Kitchen andbathroom walls, ceilings, and fixtures attached thereto, as well asshelving, cabinetry, are examples of surfaces to which to apply thesolutions and fluid compositions of the present invention. Similarly inthe kitchen and laundry, ceilings and walls and the fixtures attachedthereto, counter tops, open and closed shelving, cabinetry, stoves,refrigerators, washing machines, drying machines, microwave ovens, andfurniture are examples of objects with surfaces to which to applysolutions and fluid compositions of the present invention.

Similarly in places of business, government, and public education thereare walls and ceilings, fixtures attached thereto, banisters, doors,windows, furniture, lockers, rest rooms, auditoriums, vestibules,cafeteria and snack shop areas, and other surfaces and substrates towhich the solutions and fluid compositions of the present invention maybe applied.

Hospitals, medical centers, doctor's offices, and health centers are anadditional class of places that comprise walls, ceilings, fixturesattached thereto, operating rooms, waiting rooms, rest rooms, bathrooms, laboratories, hallways, nurse's stations, patient rooms, windows,window treatments, diverse curtains, doors, cabinetry and diverseappliances, machinery, and equipment that exhibit surfaces andsubstrates to which the solutions and fluid compositions of the presentinvention may be applied.

Certain substrates of the present invention are themselves liquidsuspensions and solutions. Topical creams for treating wounds, yeastinfections, acne, superficial burns are suitable substrates to which thesolutions, fluids, and compounds of the present invention may beusefully applied.

Polymer Binders and Resins

The compounds of the present invention may be suitably incorporated intoa microemulsion of immiscible monomers and form polymeric coatings bymicroemulsion polymerization, as described in Texter et al. (2004)Macromolecules 37: 5841-5843; (2005) Correction 37: 7424, herebyincorporated by reference. Microemulsion polymerization is a method forformulating polymer coatings from immiscible monomers. Microemulsionsare thermodynamically stable solutions of at least two immisciblefluids, such as oil and water, made miscible by the action of a thirdchemical component, suitably a surfactant such as silverbis[2-ethylhexyl]sulfosuccinate or sodiumbis[2-ethylhexyl]sulfosuccinate. For example, a polyurethane coating maybe made from a microemulsion of propylene glycol and isophoronediisocyanate fluids. The microemulsion may contain equivalent weights oftwo different monomers, for example, propylene glycol and isophoronediisocyante, to make polyurethane. Polyurethanes may form by additionstep polymerization of diols with diisocyanates. Cross-linking agentsmay be included in the reaction to increase the molecular weight of thepolymer. A reverse microemulsion or reverse micelle may also be formed.Suitably, compounds of the invention form reverse micelles.

The polymer binders and resins of the inventions may be any of the typeincluded in commercially available paints and other protective coatingformulations, wherein the binder comprises organic or partially organicpolymeric components or wherein the binder comprises precursors topolymers, wherein such precursors transform to the final binder upondrying or curing of the paint or coating formulation.

The polymeric coatings may be hydrophobic. The polymeric binder in theform of precursors may be included in the coating composition ofbactericidal composition in the form of monomers in a microemulsion ofin the form of a solution. The microemulsion may further include atleast about 5% to at least about 50% of silver anionic surfactant byweight relative to the weight of all monomers and anionic surfactant.The microemulsion may include less than about 95% to less than about 50%silver or sodium anionic surfactant. Suitably, the microemulsionincludes about 0.01 to 3% silver anionic surfactant by weight relativeto the weight of all monomers and anionic surfactant. The microemulsionmay also include more than one cationic form of the anionic surfactant.For example, a 30% anionic surfactant microemulsion may contain 10%silver anionic surfactant and 20% sodium anionic surfactant, or 1%silver anionic surfactant and 29% sodium anionic surfactant, or 0.1%silver anionic surfactant and 29.9% sodium anionic surfactant. Themicroemulsion is suitably placed on a surface and thereon cured byheating or by UV irradiation to form a polymeric coating on the surface.Suitable surface materials include ceramics, glass, metal such asaluminum and steel, plastics, and wood. Suitable surfaces and substratesinclude any material in the consumer, industrial, and government arenathat benefits from a decorative or protective coating. A catalyst may beadded to the microemulsion to drive polymerization on a surface.Suitable catalysts include dibutyltin dilaurate (DBTD).

The polymeric coatings in accordance with the invention may be at leastabout 1 μm thick to at least about 500 μm thick as needed for theprotective application. Suitably, the polymeric coating may be about 5to 50 μm thick. The polymeric composition in accordance with theinvention may contain about 0.0001% to about 1.0% metal cation, suitablyabout 0.0001% to about 1.0% Ag⁺, by weight of the total composition.

Polymeric Compositions

Polymer compositions of the invention include a plurality ofthermoplastic polymers that may be used to form objects useful in theconsumer, industrial, and healthcare industries. Such objects may bemade by any appropriate means of fabrication, including extrusion,injection molding, blow molding, casting, and solvent casting. Suchobjects may include bottles for storing foods, liquids, cosmetics, andperishable items. They may also include polymers to be fabricated tocatheters and tubing, artificial joints, bags, and packaging materials.In these types of compositions, the silver compounds of the inventionmay be incorporated at appropriate levels, determined by experimentationillustrated in the examples, during the polymer formation process.Alternatively, the silver compounds of the invention may be incorporatedby swelling the polymer in a suitable solvent containing the silvercompounds, and allowing the object to dry. This process may provide agradient of active silver in the materials that is highest at thesurface and decreases with depth of penetration into the surface.

The silver containing compositions, coatings, and solutions of theinvention inhibit growth or activity of microorganisms or microbes,including bacteria. The compositions inhibit growth or activity ofmicroorganisms or microbes, including bacteria. The silver surfactantcompounds and compositions of the invention suitably reduce theviability of Gram-negative bacteria including Escherichia coli;Salmonella; Pseudomonas such as Pseudomonas aeruginosa; Proteusmirabilis; Enterobacter cloacae; Enterobacteria aerogenes; Serratiamarcescens; Neisseria such as Neisseria gonorrhoeae and Neisseriameningitides; Moraxella catarrhalis; Hemophilus influenzae; Klebsiellapneumoniae; Legionella pneumophila; Helicobacter such as Helicobacterpylori; Salmonella such as Salmonella enteritidis and Salmonella typhiand Salmonella typhimurium; and Acinetobacter baumanii. The sodiumsurfactant compounds and compositions of the invention suitably reducethe viability of Gram-positive bacteria including Bacillus such asBacillus anthracis, Bacillus cereus, Bacillus subtilis; Listeria;Staphylococcus such as Staphylococcus aureous and Staphylococcusepidermidis; Streptococcus such as Streptococcus viridans andStreptococcus bovis and Streptococcus pyogenes and Streptococcusagalactiae; Enterococcus such as Enterococcus fecalis; Clostridium; andMicrococcus luteus. The sodium surfactant compounds and compositions ofthe invention may also reduce the viability of yeast such as Candidatropicalis.

Without being held to any particular theory, antibacterial compounds ofthe invention may act by disrupting cell membranes, such as by inducinglysis or affecting cell transport. Silver ion may, for example, bind tothiols in enzymes and proteins to deactivate the enzymes and proteins.Silver ions may also interfere with ion transport and respiration acrossa cytoplasmic membrane. Activity of the surfactant compounds asantimicrobial agents may also be related to the tendancy of the compoundto partition the bacterial bilayer membrane with concomitant disruptionof membrane function.

Silver containing compositions, coatings, and solutions of the inventionmay offer environmentally friendly approaches to guarding againstGram-positive and Gram-negative bacteria in coating and cleaningformulations. Compositions of the invention may also combat antibioticresistant staphylococcus and pneumonia coccus infections. Compounds ofthe invention may be used to make materials that are antimicrobial.Compounds of the invention may be incorporated or impregnated into, orapplied to medical devices or other articles of manufacture such ascatheter materials, syringes, wound dressings. Compounds of theinvention may be incorporated into antimicrobial or prophylacticformulations such as coating formulations, sprays, medical treatmentsfor burns or infections, wipes or swabs for treating acne or preventingor reducing infection, coating compositions for medical devices. Thecompounds and compositions of the invention may be applied to substratesincluding mammalian tissue, wound dressing materials, fabrics, syntheticabsorbent materials, clothing articles, ceilings, walls, wall fixtures,ceiling fixtures, window treatments, shelving, fixed and moveablefurniture, banisters, appliances, computers, machines, and filingcabinets

EXAMPLES

The following non-limiting examples are purely illustrative.

Example 1 Preparation of silver bis[2-ethylhexyl]sulfosuccinate byColumn Ion Exchange

The sodium salt of bis[2-ethylhexyl]sulfosuccinate was obtained fromFisher Scientific (Hanover, Ill., U.S.A.). The silver form of thecompound, silver bis[2-ethylhexyl]sulfosuccinate, was generated by ionexchange of sodium bis[2-ethylhexyl]sulfosuccinate over the strongcationic resin LewatitSP 1080 (LANXESS, Pittsburgh, Pa., U.S.A.) thatwas previously protonated. The acid form of the compound was then passedthrough the weakly acidic macroporous cation exchange resin Bio-Rex 70resin (BIO-RAD, Hercules, Calif., U.S.A.) after it had been loaded withsilver ion. The solvent used for both ion exchange columns was 50/50ethanol-water solution. The resulting silver form ofbis[2-ethylhexyl]sulfosuccinate was dried under vacuum and stored in adessicator until used.

Example 2 Preparation of silver bis[2-ethylhexyl]sulfosuccinate by TwoPhase Ion Exchange

Sodium bis(2-ethylhexyl)sulfosuccinate (NaAOT) is obtained from Fisher.10 g NaAOT is dissolved in 100 mL diethylether and combined with anequal volume of 0.1 M aqueous AgNO₃ in a separatory flask. The mixtureis shaken and the aqueous phase was decanted. Two additional 100 mLvolumes of 0.1 M AgNO₃ are similarly ion exchanged with the ethersolution. The ether solution is then washed twice with deionized water.The aqueous phase is decanted and the ether phase is reduced to a pasteusing rotary evaporation. The remaining paste is dried in a vacuum ovenat 25-50° C., and a yield in excess of 90% is obtained.

Example 3 Preparation of silver dodecylsulfate

Silver dodecylsufate (AgDS) is made by precipitating an aqueous solutionof sodium dodecylsulfate with a 5% excess of silver nitrate. Theprecipitate is washed with water and then recrystallized from hot water.

Example 4 Preparation of silver linearalkanebenzene sulfonate

Acid linearalkanesulfonate is obtained from Aldrich and precipitatedfrom aqueous solution with 5% excess silver nitrate. The precipitate isfiltered and washed with water, and then recrystallized from hot water.

Example 5 Preparation of silver bis[2-ethylhexyl]phosphate by Reactionwith silver oxide

Acid bis(2-ethylhexyl)phosphate is obtained commercially fromSigma-Aldrich. The liquid acid, in methanol, is reacted with anequivalent of silver oxide. Quantitative conversion is obtained. Thesilver bis(2-ethylhexyl)phosphate (AgDEHP) is dried in vacuo.

Example 6 Preparation of silver poly(AMPS-co-EHA) by Reaction withsilver oxide

2-Acrylamido-2-methyl-1-propanesulfonic acid (AMPS) and 2-ethylhexylacrylate are obtained from Sigma-Aldrich (St. Louis, Mo., U.S.A.) andthe two monomers are copolymerized in THF/DMF at 10% total monomer at60° C. using AIBN as initiator at 0.5% by weight relative to the totalmonomers. After polymerization 5% water is added to the product solutionand then this solution is reacted with an equivalent amount of silveroxide with vigorous stirring. The reaction is reduced in volume at 60°C. by rotary evaporation, followed by drying in a vacuum oven overnight.

Example 7 Preparation of poly(styrene-co-styrenesulfonate, silver salt)by Reaction with silver oxide

Polystyrene (400,000 g/mol) is sulfonated with acetylsulfate asdescribed by Makowski, Lundberg, and Sighal in U.S. Pat. No. 3,870,841and by W. Chen, J. A. Sauer, and M. Hara (Polymer 45 (2004) 7219-7227).After the sulfonation reaction is terminated by the addition ofmethanol, polymer (acid form) is recovered by steam stripping in boilingwater. After air drying for 3 days, the polymer is dissolved in asolvent mixture, toluene/methanol (90:10 v/v), and freeze-dried,followed by vacuum drying at room temperature for a week. The acidcontent was determined by potentiometric titration with sodium methoxidein methanol to be 3.7 mole %. The polymer is redissolved intoluene/methanol/water (80/15/5) and an equivalent of powdered silveroxide is added. The reaction is stirred overnight, and then dried on arotary evaporator at 50° C. followed by drying in a vacuum oven at roomtemperature. Analysis for silver indicates complete conversion with 52mg silver ion per g polymer.

Example 8 Preparation of poly(styrene-co-styrenesulfonate, silver salt)by Two-Phase Ion Exchange

The acid sulfonate form of poly(styrene-co-styrenesulfonate) describedin Example 7 is titrated to the endpoint with sodium methoxide (1.0 M inmethanol), and the sodium salt is dissolved in toluene. This solution isplaced with an equal volume of 0.1 M silver nitrate in a separatoryflask, shaken, and the lower aqueous phase decanted. This procedure isrepeated twice more, and then the toluene phase is washed twice withdeionized water. The toluene solution is reduced in volume by rotaryevaporation, and the remaining paste is dried in a vacuum oven at 40° C.overnight.

Example 9 Preparation of 0.06% (w/w) silver Solution of silverbis[2-ethylhexyl]sulfosuccinate in isopropanol

The silver bis(2-ethylhexyl)sulfosuccinate described in Example 2 isdissolved in isopropanol to prepare a solution 0.06% (w/w) silver inisopropanol by combining 290 mg of the silver surfactant with 99.71 g ofisopropanol.

Example 10 Preparation of 0.24% (w/w) silver Solution of silverbis[2-ethylhexyl]sulfosuccinate in isopropanol

The silver bis(2-ethylhexyl)sulfosuccinate described in Example 2 isdissolved in isopropanol to prepare a solution 0.24% (w/w) silver inisopropanol by combining 1.16 g of the silver surfactant with 98.84 g ofisopropanol.

Example 11 Preparation of 0.10% (w/w) silver Solution ofpoly(styrene-co-styrenesulfonate, silver salt)

The silver sulfonate form of poly(styrene-co-styrenesulfonate) describedin Example 7, 192 mg, is dissolved in 9.81 g of toluene to yield asolution 0.10% w/w in silver ion.

Example 12 Preparation of Bacterial Broth Cultures

Escherichia coli ATCC 11229 (Culti-loops, Remel Europe, Dartford, Kent,UK) and Pseudomonas aeruginosa ATCC 15442 (Cuti-loops, Remel Europe,Dartford, Kent, UK) were selected as model Gram-negative organisms forthe challenge testing of the antimicrobial coatings. Each bacterialstrain was streaked for isolated colonies on Tryptic Soy Agar (TSA)plates (Difco, Becton Dickinson, Sparks, Md., U.S.A.) and incubated at37° C. for 12 to 18 hours. Broth cultures were prepared by first picking1-2 isolated colonies from TSA plates and inoculating 5 mL Tryptic SoyBroth (TSB; Difco, Becton Dickinson, Sparks, Md., U.S.A.) in a 16 mmtest tube. This culture was grown in a shaking water bath at 150 RPM for8 hours at 37° C. After the eight hours, 125 μL of this culture wasadded to 50 mL of TSB in a side-arm Klett flask and grown forapproximately 8 hours in shaking water bath (150 RPM) at 37° C. Thebacterial broths were grown to an A₅₉₀ of 1.2-1.5, which correlated toapproximately 109 CFU (colony forming units)/mL. 25 mL of this brothwere transferred to a sterile 30 mL Oakridge tube and centrifuged at12,000×g for 10 minutes at 4° C. (Sorval SS34 rotor) to pelletize thebacteria. The bacterial pellet was washed once and resuspended in 25 mLof phosphate buffer solution (PBS; BBL, FTA Hemaglutination Buffer,Becton Dickinson, Sparks, Md., U.S.A.). The suspension was diluted withPBS to achieve the target range of 105-106 CFU/mL and used to inoculatethe slides containing antibacterial coatings.

Example 13 Preparation of a Bacterial Cultures and Coupon TestingProtocol

The antimicrobial activities of the coatings on coupons and slides aretested against both Gram-positive (Staphylococcus aureus ATCC 6538) andGram-negative (Escherichia coli ATCC 11229) bacteria. The following setof steps are executed over a four day period, in order to evaluate thecoatings for antimicrobial activity.

A. Overnight Culture Bacterial Cultures (Day 1)

(1) Three colonies are picked from a Petri dish that containsrefrigerator stock of E. coli and S. aureus. Each of the colonies isplaced into a separate test tube of 5 mL of TSB (tryptic soy broth); (2)the TSB cultures are incubated in a heated shaker; the culture is grownovernight at 37° C. while shaking at 175 rpm.

B. Grow Cultures for 3-4 Hours (Day Two)

(1) Three Klett flasks are sterilized by autoclaving (gravity cycle for15 minutes and dry for 15 min); (2) 50 mL of TSB is added to each flask;(3) 5 mL of E. coli overnight culture is added to one flask, 5 mL of S.aureus overnight culture is added to the second, while the third is leftas a control for absorbance readings; (4) the cultures are grown for 3to 4 hours with shaking at 150 rpm and at 37° C.; (5) the absorbance at600 nm is checked to confirm the transmittance read 0.85-0.9 to ensurethe bacteria was 1.2-1.5×10⁹ CFU/mL.

C. Preparing Petri Dishes (Day 2)

(1) The empty Petri dishes are labeled and coupons or slides are placedin them, with coating side facing down first; (2) Petri dishes aresterilized under UV light for 2 minutes on each side; slides or couponsare flipped with sterile forceps after the initial 2 minutes; prior tothe testing the slides are sterilized by exposure to a germicidal UVlamp in a Nuaire Model NU-455-600 Class II Type A/B3 laminar flow hood(2 min on each side at room temperature); (3) Tryptic Soy Agar (TSA)plates are labeled as 0/24 hour, SA/EC, coupon #, and concentration ofinoculums to be added (−1,−2,−3); two plates are made for eachconcentration and for each coupon for a total of six plates; (4) TSAplates are labeled as inoculums for each type of bacteria (SA/EC), at 0hour, and concentrations of −3,−4,−5; 2 of each are made.

D. Centrifuge (Day 2)

(1) Once cultures have grown for 3-4 hours, the absorbance is checkedand when adequate (1.2-1.5 ×10⁹ CFU/mL), 25 mL are transferred toplastic centrifuge tubes and centrifuged for 10 min. @ 10,000 rpm at 4°C. in Sorval SS34 rotor; (2) the tubes are removed from centrifuge andthe TSB is poured out, leaving only the pellet of bacteria in the bottomof tube; immediately 25 mL of phosphate buffer solution (PBS) is addedto the remaining pellet of bacteria and the pellet i resuspended byshaking with vortex mixer; (3) two new plastic tubes (conical tubes) arelabeled for each type of bacteria, and labeled as 1st and FINAL; (4) 9.9mL of PBS is added to 0.1 mL (100 microliters) of the above preparedsolution (step 2) into 1st tube; the concentration of the bacteria ischanged from 10⁹ to 10⁷ CFU/mL; (5) 9.9 mL of PBS was diluted into FINALtube by adding 0.1 mL (100 microliters) of contents from the 1st tube;the concentration of bacteria is diluted from 10⁷ to 10⁵ CFU/mL; this isthe FINAL bacteria used to inoculate the coating being evaluated.

E. Inoculation of Sterilized Coupons—24 Hour Coatings (Day 2)

(1) 500 μL (0.5 mL) of FINAL E. coli bacteria are placed onto the centerof each sterilized coating (24 hour); the same is done with S. aureus;(2) the coupons/coatings are stored for 24 hours in sealed pan bystacking Petri dishes on top of one another; (3) 200 mL of sterile wateris added to the bottom of each pan; each pan is enclosed with clearsaran wrap and aluminum foil; the Pyrex pans containing the coatings areincubated at room temperature for 24 hours; all inoculations of thecoatings are performed in the same laminar flow hood as where thesterilized slides are placed into sterile Petri dishes; the preparedbacterial suspension (0.5 ml) is directly pipetted onto the coating ofthe slide; the spreading of the innoculum across the surface of thecoating is observed and noted.

F. Inoculation of Sterilized Coupons—Zero Hour Coatings (Day 2)

(1) 500 μL (0.5 mL) of FINAL E. coli bacteria is placed onto the centerof each sterilized coating (0 hour); the same is done with S. aureus;(2) after bacteria are added to the 0 hour coatings, each Petri dish(containing the coating) is submerged with 25 mL of PBS; this immersionstops the viable bacteria from continuing to grow; (3) 100 μL of fluidis removed from coatings dish and placed in the middle of a previouslylabeled TSA plate; this is done to both agar plates labeled with −1concentration; before removing the fluid, the surface of the coupon isscratched with a sterile loop to activate movement of bacteria; (4)“hockey sticks” are sterilized with ethanol and flame, to spreadsolution thoroughly; (5) the concentration of bacteria is diluted byremoving 500 μL of the fluid in the coupon dish and placing it into atest tube with 4.5 mL of PBS; this solution is diluted in duplicate byremoving 100 μL of the fluid and spreading into TSA plates labeled −2;(6) sterilized “hockey sticks” are used to spread solution thoroughly;(7) 500 μL of fluid from 1st tube prepared in step 5 is removed andplaced into 4.5 mL of PBS; this dilution is mixed and 100 μL is removedin duplicate and placed into two new TSA plates labeled −3; (8)sterilized “hockey sticks” are used to spread these dilutionsthoroughly; (9) steps 1-8 are repeated for each of the coupons orslides.

G. Control Inoculum (Day 2)

(1) 500 μL of solution from coupon dish labeled control (blank glassslide) are removed and added to 4.5 mL of PBS; this dilution is mixedand 500 μL of this solution is removed and added to another 4.5 mL ofPBS in a test tube; this dilution is mixed and 100 μL is removed andplaced in control SA or EC TSA plates labeled −3; (2) these dilutionsare spread thoroughly; (3) another dilution is prepared by removing 500μL of the solution in the test tube with the −3 concentration and addingto a new 4.5 mL of PBS in a Petri dish; this dilution is mixed and thedish labeled −4 concentration; (4) step 3 is repeated for a platelabeled −5; (5) all TSA plates are incubated overnight in a 37° C. oven.

H. Recovery of Surviving Bacteria (Days 3 and 4)

After 24 hours, TSA plates (incubated from the previous day) containingbetween 20 and 300 isolated colonies are counted. The coupons that hadbeen stored in Pyrex pans for 24 hours are treated as the coupons instep F (above) are treated. Again, these plates are incubated overnightin a 37° C. oven. After twenty-four hours, surviving colonies arecounted and recorded.

Example 14 Spray Application of silver bis(2-etylhexy)sulfosuccinate toFabrics

Fabric samples are obtained commercially of cotton, linen, polyester,acetate, rayon, and silk. The 0.06% silver ion solutions described inExample 9 are used. Application of solutions is done using an art-supplyairbrush gun; air is supplied from a house compressed supply, under aventilated hood. Fabric is carefully masked so no spray was lost. Allspray impact is kept within a 5 cm×8 cm masked surface. Afterapplication the fabrics are allowed to hang and air dry for 18 hours.The fabrics are then cut into 10 cm² (2 cm×5 cm) pieces beforesterilizing. The fabric swatches are then placed in large glass Petridishes, avoiding contamination after removing from autoclave. Fabricswatches coated with AgAOT are autoclaved (sterilized) at 121° C. (18psi, gravity cycle, 30 minutes of sterilization, and 5 minutes of drytime). The AgAOT-coated fabrics all turn a brownish color, oncesterilized at 121° C.

TABLE 4 S. aureus challenge with 0.06% w/w AgAOT/isopropanol; CFU/mL at0 h contact time. Fabric Run #1 Run #2 Run #3 Run #4 Average Std DevInoculum 2.50E+05 2.30E+05 4.50E+05 3.00E+05 3.08E+05 9.95E+04 Cottonw/iso 2.80E+05 2.10E+05 2.50E+05 2.20E+05 2.40E+05 3.16E+04 w/0.06% Ag+0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Linen w/iso1.20E+05 2.20E+05 8.30E+04 7.40E+04 1.24E+05 6.69E+04 w/0.06% Ag+0.00E+00 4.80E+04 2.50E+02 1.50E+03 1.24E+04 2.37E+04 Polyester w/iso1.70E+07 1.80E+05 1.60E+05 1.50E+05 4.37E+06 8.42E+06 w/0.06% Ag+0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Acetate w/iso2.70E+05 2.30E+05 3.20E+05 2.00E+05 2.55E+05 5.20E+04 w/0.06% Ag+0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Rayon w/iso1.70E+05 2.30E+05 3.20E+05 3.30E+05 2.63E+05 7.63E+04 w/0.06% Ag+0.00E+00 0.00E+00 2.50E+02 5.00E+02 1.88E+02 2.39E+02 Silk w/iso1.30E+05 1.90E+05 1.60E+05 4.20E+04 w/0.06% Ag+ 1.80E+05 1.10E+051.45E+05 4.90E+04

Example 15 S. aureus Challenge by Fabrics Sprayed with AgAOT; 0 hContact Time

The test fabric swatches are then used to challenge S. aureus asdescribed in Example 13 with two exceptions: (1) conical flasks are usedto hold fabrics overnight, not Petri dishes; (2) only 5 mL of PBS isused to stop the growth of bacteria, instead of 25 mL of PBS. Thechallenge results for 0 h contact time are presented in Table 4.

The results show that the level of silverbis(2-ethylhexyl)sulfosuccinate coated on each of the fabricssignificantly arrests the S. aureus growth in each of the fabricstested, except for silk. The growth inhibition is complete in the casesof cotton, polyester, and acetate. Inhibition on rayon is by 3 orders ofmagnitude. That on the linen is only by one order of magnitude, andthere appears to be no effect for silk. Since the 0 h contact time inreality is about 3-10 minutes, the inhibition that occurs suggests theinhibition activity or bactericidal activity of the invention silversurfactant is significant.

TABLE 5 S. aureus challenge with 0.06% w/w AgAOT/isopropanol; CFU/mL at24 h contact time. Fabric Run #1 Run #2 Run #3 Run #4 Average Std DevCotton Blank 1.00E+05 6.00E+04 6.40E+04 6.10E+04 7.13E+04 1.92E+04 w/iso6.30E+04 1.00E+05 5.10E+04 1.10E+05 8.10E+04 2.84E+04 w/0.06% Ag+0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Linen Blank1.00E+04 1.00E+04 3.80E+04 1.90E+04 1.93E+04 1.32E+04 w/iso 1.00E+045.00E+03 1.30E+04 4.80E+03 8.20E+03 4.00E+03 w/0.06% Ag+ 0.00E+000.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Polyester Blank 1.50E+051.10E+05 2.30E+05 2.00E+05 1.73E+05 5.32E+04 w/iso 1.20E+05 9.30E+041.90E+05 1.00E+05 1.26E+05 4.43E+04 w/0.06% Ag+ 0.00E+00 0.00E+000.00E+00 0.00E+00 0.00E+00 0.00E+00 Acetate Blank 2.20E+05 1.30E+051.10E+05 2.80E+05 1.85E+05 7.94E+04 w/iso 1.30E+05 1.90E+05 2.30E+051.40E+05 1.73E+05 4.65E+04 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+000.00E+00 0.00E+00 0.00E+00 Rayon Blank 2.00E+05 1.20E+05 9.80E+041.70E+05 1.47E+05 4.64E+04 w/iso 2.30E+05 1.70E+05 1.30E+05 8.80E+041.55E+05 6.05E+04 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+000.00E+00 0.00E+00 Silk Blank 7.10E+06 CG 7.10E+06 w/iso 1.80E+061.30E+06 1.55E+06 3.50E+05 w/0.06% AgAOT 2.50E+02 1.80E+04 9.13E+031.26E+04

Example 16 S. aureus Challenge by Fabrics Sprayed with AgAOT; 24 hContact Time

The test fabric swatches are then used to challenge S. aureus asdescribed in Examples 13 and 15, except the contact time is 24 h. Theresults are presented in Table 5.

The results show that the level of silverbis(2-ethylhexyl)sulfosuccinate coated on each of the fabricssignificantly arrests the S. aureus growth in each of the fabricstested. It appears that complete eradication is obtained in the cases ofcotton, linen, polyester, acetate, and rayon. The growth on silk isattenuated by two orders of magnitude compared to the controls, but thekilling is incomplete, and confluent growth (CG) is observed on the silkblank sample.

Example 17 E. coli Challenge by Fabrics Sprayed with AgAOT; 0 h ContactTime

The test fabric swatches are then used to challenge E. coli as describedin Examples 13 and 15 for a contact time of 0 h. The results arepresented in Table 6. The short 0 h contact time (5-10 minutes) do notsignificantly attenuate E. coli growth. Some partial attenuation isapparent in the cases of linen and polyester, however.

TABLE 6 E. coli challenge with 0.06% w/w AgAOT/isopropanol; CFU/mL at 0h contact time. Fabrics Run #1 Run #2 Run #3 Run #4 Average Std DevInoculum 3.60E+05 2.40E+05 4.50E+05 3.00E+05 3.38E+05 8.96E+04 Cottonw/iso 3.50E+05 1.90E+05 2.50E+05 3.20E+05 2.78E+05 7.18E+04 w/0.06% Ag+3.00E+05 2.50E+05 2.60E+05 2.00E+05 2.53E+05 4.11E+04 Linen w/iso1.50E+05 1.50E+05 8.30E+04 3.10E+05 1.73E+05 9.65E+04 w/0.06% Ag+2.60E+05 2.60E+05 2.50E+02 6.30E+04 1.46E+05 1.34E+05 Polyester w/iso2.20E+05 2.70E+05 1.60E+05 3.20E+05 2.43E+05 6.85E+04 w/0.06% Ag+3.60E+05 5.00E+02 0.00E+00 3.30E+03 9.10E+04 1.79E+05 Acetate w/iso2.80E+05 2.70E+05 1.60E+05 2.50E+05 2.40E+05 5.48E+04 w/0.06% Ag+3.80E+05 2.60E+05 3.60E+05 2.30E+05 3.08E+05 7.37E+04 Rayon w/iso2.40E+05 2.30E+05 4.50E+05 3.70E+05 3.23E+05 1.06E+05 w/0.06% Ag+3.20E+05 2.10E+05 3.50E+05 2.70E+05 2.88E+05 6.13E+04 Silk w/iso2.50E+05 1.50E+05 2.00E+05 7.00E+04 w/0.06% Ag+ 2.70E+05 2.40E+052.55E+05 2.10E+04

Example 18 E. coli Challenge by Fabrics Sprayed with AgAOT; 24 h ContactTime

The test fabric swatches are then used to challenge E. coli as describedin Examples 13 and 15 for a contact time of 24 h. The results arepresented in Table 7. With the single exception of silk, completeeradication is obtained for each of the other fabrics by the appliedsilver bis(2-ethylhexyl)sulfosuccinate.

Example 19 Silver Binding by Silk

The weave of the silk samples was investigated by scanning electronmicroscopy, and the woven fabric appeared very similar to the othernatural and synthetic weaves. Further, no dewetting of the appliedinvention solution was observed after careful observation. The silverion binding to the silk swatches of these Examples are quantitativelyinvestigated.

TABLE 7 E. coli challenge with 0.06% w/w AgAOT/isopropanol; CFU/mL at 24h contact time. Fabric Run #1 Run #2 Run #3 Run #4 Average Std DevCotton Blank 1.00E+07 1.00E+07 1.00E+07 CG 1.00E+07 0.00E+00 w/iso5.00E+06 1.00E+07 1.00E+07 CG 8.33E+06 2.89E+06 w/0.06% Ag+ 0.00E+000.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Linen Blank 1.50E+071.50E+07 1.00E+07 CG 1.33E+07 2.89E+06 w/iso 1.50E+07 1.50E+07 1.00E+07CG 1.33E+07 2.89E+06 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+000.00E+00 0.00E+00 Polyester Blank 1.00E+07 1.50E+07 1.00E+07 CG 1.17E+072.89E+06 w/iso 5.00E+06 1.50E+07 1.00E+07 CG 1.00E+07 5.00E+06 w/0.06%Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Acetate Blank1.00E+07 1.70E+05 4.40E+05 3.40E+05 2.74E+06 4.84E+06 w/iso 3.30E+052.00E+05 3.90E+05 2.90E+05 3.03E+05 7.97E+04 w/0.06% Ag+ 0.00E+000.00E+00 0.00E+00 0.00E+00 0.00E+00 0.00E+00 Rayon Blank 1.00E+071.50E+07 1.50E+07 CG 1.33E+07 2.89E+06 w/iso 1.00E+07 1.50E+07 1.50E+07CG 1.33E+07 2.89E+06 w/0.06% Ag+ 1.30E+06 2.50E+02 0.00E+00 0.00E+003.25E+05 6.50E+05 Silk Blank 1.50E+07 1.50E+07 1.50E+07 w/iso 1.50E+071.50E+07 1.50E+07 w/0.06% Ag+ 5.00E+07 5.00E+06 2.75E+07 3.15E+07

Silk swatches of 2 cm×5 cm (10 cm²) are suspended in water and thesilver ion potential is recorded using a silver wire as indicatingelectrode and a AgCl reference electrode with a potassium nitrate slatbridge. Potential readings in mV are measured with a pH meter as 0.001 Msilver nitrate is titrated into the stirred solution in which the swatchis suspended. Several possible endpoints are evident, but the highestpotential one corresponds to the total silver ion binding end point, andthis is observed at 3.7 mL of added titrant. This endpoint correspondedto 0.37 μmol Ag⁺/cm². Other sprayed silk swatches are carefully weighedbefore and after the 0.06% w/w silver bis(2-ethylhexyl)sulfosuccinatesolutions are applied by spraying, as detailed in Example 14. The weightof solution applied to each swatch samples is 1.56 g. From the area ofthe total swatch and the solution composition, the coverage of 0.21 μmolAg⁺/cm² is calculated for a 0.06% w/w silver ion solution. It is evidentthat the silk fabric tightly binds 0.37/0.21=1.8 times the amount of Ag⁺applied. The negative to marginal results seen in Examples 15-18 forsilk, therefore can be understood in view of the fact that the silkitself strongly sequesters almost twice the amount of Ag⁺ applied.

Example 20 Spray Application of silver bis(2-etylhexy)sulfosuccinate toSilk

The 0.24% silver ion solutions described in Example 10 are used.Application of solutions is done as described in Example 14. This levelof coverage corresponds to 0.81 μmol Ag⁺/cm2, over twice the amount ofsilver ion sequestered by a 2 cm×5 cm swatch of silk.

Example 21 S. aureus Challenge by Fabrics Sprayed with AgAOT; 0 hContact Time

The swatches of Example 20 were tested as described in Examples 15-18 inchallenging both S. aureus and E. coli for 0 and 24 h contact times. Theresults in Table 8 show that while neither organism was significantlyaffected at 0 h contact time (5-10 minutes), both were eradicatedcompletely after 24 h contact.

Example 22 Spray Application of silver bis(2-ethylhexy)sulfosuccinate toHousehold Surfaces

Three samples of household interior surfaces are obtained from ahardware store. “Metal plate” is a painted metal cover plate for aconventional in-wall mounted light switch. “Wall paper” is a sample ofwall paper used for interior wall decoration. “Vinyl tile” is a sampleof vinyl floor covering used to coat interior kitchen floors andcounters. The Metal plate and Vinyl tile samples are cut into 2 cm×5 cmsamples; two sets of four samples are arranged in an 8 cm×5 cm area forspray application of isopropanol (one set) and for spray application ofa 0.06% silver solution described in Example 9. The Wall paper samplesare cut into 5 cm×8 cm rectangles; one is sprayed with isopropanol andanother is sprayed with a 0.06% silver solution described in Example 9.Application of solutions is done using an art-supply airbrush gun; airis supplied from a house compressed supply, under a ventilated hood. Thesamples are left to dry over night.

TABLE 8 S. aureus and E. coli challenges with 0.24% w/wAgAOT/isopropanol on silk; CFU/mL. Treatment Run #1 Run #2 Run #3 Run #4Average Std Dev S. aureus - 0 h w/iso 1.90E+05 w/0.24% Ag+ 9.00E+041.40E+05 1.70E+05 1.40E+05 1.35E+05 3.32E+04 S. aureus - 24 h Blank7.10E+06 CG w/iso 2.30E+06 1.70E+06 w/0.24% AgAOT 0.00E+00 0.00E+000.00E+00 0.00E+00 0.00E+00 0.00E+00 E. coli - 0 h w/iso 3.50E+05 w/0.24%Ag+ 3.20E+05 3.60E+05 3.60E+05 3.10E+05 3.38E+05 2.63E+04 E. coli - 24 hBlank CG CG w/iso CG CG w/0.24% Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+000.00E+00 0.00E+00

The Wall paper samples are then cut into 10 cm² (2 cm×5 cm) piecesbefore sterilizing. The 5 cm×2 cm samples are then placed in large glassPetri dishes, avoiding contamination after removing from autoclave. Thesamples coated with AgAOT are autoclaved (sterilized) at 121° C. (18psi, gravity cycle, 30 minutes of sterilization, and 5 minutes of drytime).

Example 23 S. aureus Challenge by Surfaces Sprayed with AgAOT; 0 hContact Time

The household surface samples described in Example 22 are then used tochallenge S. aureus for 0 h contact time, as described in Example 13with two exceptions: (1) conical flasks are used to hold fabricsovernight, not Petri dishes; (2) only 5 ml of PBS are used to stop thegrowth of bacteria, instead of 25 ml of PBS. The challenge results for 0h contact time are presented in Table 9.

TABLE 9 S. aureus challenge by surfaces sprayed with AgAOT; CFU/mL at 0h contact time. Surface/Substrate Run #1 Run #2 Avg Std Dev Inoculum1.50E+05 1.80E+05 1.65E+05 0.21E+05 Metal plate w/iso 9.60E+05 2.20E+055.90E+05 5.18E+05 w/0.06% Ag+ 0.00E+00 7.90E+04 3.90E+04 5.46E+04Wall-paper w/iso 9.60E+05 2.40E+05 6.00E+05 5.04E+05 w/0.06% Ag+5.00E+04 1.90E+05 9.75E+04 6.65E+04 Vinyl tile w/iso 7.20E+05 1.90E+054.55E+05 3.71E+05 w/0.06% Ag+ 1.50E+02 2.50E+05 1.25E+05 1.75E+05

Example 24 S. aureus Challenge by Surfaces Sprayed with AgAOT; 24 hContact Time

The household surface samples described in Example 22 are then used tochallenge S. aureus as described in Example 23, except the contact timeis 24 h. The results are presented in Table 10. Complete killing isobtained for each of the surfaces with the invention silverbis(2-ethylhexyl)sulfosuccinate solution.

TABLE 10 S. aureus challenge by surfaces sprayed with AgAOT; CFU/mL at24 h contact time. Surface/Substrate Run #1 Run #2 Avg Std Dev Metalplate Blank 2.70E+05 5.60E+04 1.63E+05 1.50E+05 w/iso 2.60E+05 4.20E+041.52E+05 1.51E+05 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+00Wall-paper Blank 9.20E+04 2.60E+04 5.90E+04 4.62E+04 w/iso 7.70E+043.30E+02 3.87E+04 5.36E+04 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+000.00E+00 Vinyl tile Blank 7.40E+05 2.60E+05 5.00E+05 3.36E+05 w/iso2.80E+05 4.80E+04 1.64E+05 1.62E+05 w/0.06% Ag+ 0.00E+00 0.00E+000.00E+00 0.00E+00

Example 25 E. coli Challenge by Surfaces Sprayed with AgAOT; 0 h ContactTime

The test household surface samples described in Example 22 are then usedto challenge E. coli as described in Examples 13 and 15 for a contacttime of 0 h. The results are presented in Table 11. It is clear that theshort 0 h contact time (5-10 minutes) does not significantly attenuateE. coli growth.

TABLE 11 E. coli challenge by surfaces sprayed with AgAOT; CFU/mL at 0 hcontact time. Surface/Substrate Run #1 Run #2 Avg Std Dev Inoculum3.90E+05 1.60E+05 2.75E+05 1.61E+05 Metal plate w/iso 3.90E+05 3.80E+053.85E+05 0.07E+05 w/0.06% Ag+ 3.40E+05 2.80E+05 3.10E+05 0.42E+05Wall-paper w/iso 2.90E+05 2.80E+05 2.85E+05 0.07E+05 w/0.06% Ag+8.70E+04 3.00E+05 1.94E+05 1.48E+05 Vinyl tile w/iso 3.00E+05 3.50E+053.25E+05 0.35E+05 w/0.06% Ag+ 2.30E+05 2.90E+05 2.60E+05 0.42E+05

Example 26 E. coli Challenge by Surfaces Sprayed with AgAOT; 24 hContact Time

The test household surface samples described in Example 22 are then usedto challenge E. coli as described in Examples 13 and 15 for a contacttime of 24 h. Confluent growth (CG) is observed on some of the plates.The results are presented in Table 12. E. coli is eradicated on each ofthe samples after 24 h contact time.

TABLE 12 E. coli challenge by surfaces sprayed with AgAOT; CFU/mL at 24h contact time. Surface/Substrate Run #1 Run #2 Avg Std Dev Metal plateBlank 1.30E+06 5.10E+05 9.05E+05 5.53E+05 w/iso 6.40E+05 4.10E+055.25E+05 1.61E+05 w/0.06% Ag+ 0.00E+00 0.00E+00 0.00E+00 0.00E+00Wall-paper Blank CG CG CG w/iso CG 6.40E+04 w/0.06% Ag+ 0.00E+000.00E+00 0.00E+00 0.00E+00 Vinyl tile Blank 3.80E+05 3.50E+05 3.65E+050.21E+05 w/iso 2.80E+05 8.10E+04 1.81E+05 1.40E+05 w/0.06% Ag+ 0.00E+000.00E+00 0.00E+00 0.00E+00

Example 27 Preparation of Surfactant Polymer Coated Lantern Slides

Glass lantern slides (2×3 inches, Fisher Scientific, Hanover, Ill.,U.S.A.) and microscope slides (1×3 inches, Fisher Scientific, Hanover,Ill., U.S.A.) are sterilized to rid them of any contaminatingmicroorganisms by exposing the slides to a germicidal UV lamp in aNuaire Model NU-455-600 Class II Type A/B3 laminar flow hood (2minute-exposure on each side at room temperature). The glass slides arethen washed with acetone and water.

Polyurethane coatings are formed on the glass slides in situ usingmicroemulsion polymerization as described in Texter et al. (J IndMicrobiol Biotechnol (2007) 34, 571-575). The microemulsions areformulated with equivalent weights of the immiscible step-polymerizationmonomers propylene glycol (PG) and isophorone diisocyanate (IPDI), alongwith sodium bis(2-ethylhexyl)sulfosuccinate (NaAOT) at 30% (% weightrelative to weight of all three components). Dibutyltin dilaurate (DBTD)is used as a catalyst to drive the step polymerization, and it is addedto 0.1% of the total composition weight. Coatings of approximately 150μm thickness and 5 cm width are made on glass slides using a drawdownbar. The coatings are stored for at least ten days a room temperaturebefore being subjected to challenge testing. Sodiumbis(2-ethylhexyl)sulfosuccinate coatings are used as control coatings inthe subsequent challenges. Silver bis(2-ethylhexy)sulfosuccinate(prepared as described in Example 1) is substituted for sodiumbis(2-ethylhexyl)sulfosuccinate on a weight basis from 10% to 0.01% byoverall weight, during the microemulsion formation stage prior tocoating, to produce test coatings. The 10% substitution on this weightbasis corresponds to substitution of one tenth of the total surfactantweight or to 3% of the total composition weight.

Example 28 Exposure of Bacteria to silver Surfactant (silverbis[2-ethylhexyl]sulfosuccinate) Polymer Coated Slides

Bacterial cultures as prepared and described in Example 12 are appliedto the surfactant polymer coated slides as prepared and described inExample 26. All inoculations are performed in a laminar flow hood.Sterilized slides are placed into a sterile Petri dish. Then 0.5 mL ofthe prepared bacterial suspensions is pipetted directly onto the slide,which spreads across the surface of the coating but remains on top ofthe slide. For the initial coatings provided on lantern slides andchallenged with only E. coli ATCC 11229 (2 runs), the following gradientof incorporated silver in the coatings is tested: 0%, 0.01%, 0.10%,0.50%, 1.0%, 3%, and 10% by weight of silverbis[2-ethylhexyl]sulfosuccinate is substituted for sodiumbis(2-ethylhexyl)sulfosuccinate (NaAOT) relative to the 30% controlamount of NaAOT in the coating composition on a total NaAOT weightbasis.

Example 29 Surviving Bacteria on silver Surfactant (silverbis[2-ethylhexyl]sulfosuccinate) Polymer Coated Lantern Slides

The surviving bacteria from the exposures described in Example 27 aredetermined as follows: At each time point, 25 mL of sterile PBS ispipetted into the Petri dish completely submerging the slide, and asterile plastic inoculating loop is used to release any viable bacteriaremaining on the surface of the coating. Samples are plated onto TSAplates, and incubated at 37° C. for at least 24 h. Plates containingbetween 30 and 300 isolated colonies are counted. The bacterialconcentration of the initial inoculum of bacteria used to inoculate theslides is calculated to be 4.9±1.1×10⁵ CFU/mL. The concentrations(CFU/mL) of surviving bacteria with varying concentrations of silver ionare calculated, and results are illustrated in Table 13. The data takenfrom the samples at 0 hours confirms the concentration of the inoculumused. The CFU/mL values at 24 hours demonstrates a reduction inbacterial count with increasing silver ion. In addition to the % weightof silver bis[2-ethylhexyl]sulfosuccinate relative to sodiumbis[2-ethylhexyl]sulfosuccinate in the 30% control weight gradientlisted in column 1 of Table 13, the overall weight percent of silver inthe coating composition is listed in column 2. The substitutionalpercents presented in Table 13 can be converted to overall silver weightpercents by multiplying by the factor 0.3 (control weight fraction ofsodium bis[2-ethylhexyl]sulfosuccinate), and subsequently by the factor0.204 (the weight fraction of Ag⁺ in silverbis[2-ethylhexyl]sulfosuccinate).

The results suggest that after 24 hour exposure of bacteria to thesurfactant compound, an LD₅₀ value can be inferred between thesubstitution of the AgAOT (silver bis[2-ethylhexyl]sulfosuccinate) forsodium AOT levels of 0.01% and 0.1% weight of silverbis[2-ethylhexyl]sulfosuccinate relative to 30% sodiumbis[2-ethylhexyl]sulfosuccinate control weight. Levels of substitutionat or above 0.5% weight of silver bis[2-ethylhexyl]sulfosuccinaterelative to the 30% NaAOT control weight result in complete killing ofthe E. coli. In terms of the amount of Ag⁺ in the total coatingcomposition in % weight, an LD₅₀ value can be inferred between theoverall Ag⁺ levels of 0.0006% and 0.006% weight Ag⁺ relative to coatingweight. Levels of substitution at or above 0.031% Ag⁺ relative tocoating weight result in complete killing of the E. coli.

TABLE 13 Survival of E. coli exposed to silver surfactant (silver bis[2-ethylhexyl]sulfosuccinate) polyurethane coated lantern slides. % silverbis[2- ethylhexyl]sulfosuccinate substituted for sodium bis[2-ethylhexyl]sulfosuccinate % weight silver in CFU/mL of E. coli CFU/mL ofE. coli in Coating Coating at 0 hours^(a) at 24 hours 0 0 3.7 × 10⁵; 6.2× 10⁵ 4.2 × 10⁶; 2.8 × 10⁶ 0.01 0.00061 5.4 × 10⁵; 5.5 × 10⁵ 8.4 × 10³;5.9 × 10⁶ 0.1 0.0061 4.1 × 10⁵; 4.7 × 10⁵ 1.2 × 10³; 0 0.5 0.031 4.1 ×10⁵; 4.8 × 10⁵ 0; 0 1.0 0.061 4.4 × 10⁵; 5.9 × 10⁵ 0; 0 3.0 0.18 7.5 ×10⁵; 4.7 × 10⁵ 0; 0 10.0 0.61 4.0 × 10⁵; 3.9 × 10⁵ 0; 0

Example 30 Preparation of Surfactant Polymer Coated MicroscopeSlides—Set 1

The second set of coatings was made on microscope slides and challengedwith E. coli ATCC 11229 and P. aeruginosa ATCC 15442. In these coatings,the gradient levels of incorporated silver were: 0%, 0.02%, 0.05%, 0.1%,0.2%, 0.5%, 1.0% and 2.0% of the 30% NaAOT control amount. Thesesubstitution levels are expressed as % weight of AgAOT (silverbis[2-ethylhexyl]sulfosuccinate) relative to the 30% control level ofNaAOT. These gradients were tested at three inoculation time points, 0hours, 12 hours, and 24 hours, with the 0% silver and 0 hours slides ascontrols.

The bacterial cultures were prepared as described in Example 12. Oncethe slides were inoculated, those assigned to incubate for 12 hours or24 hours were placed in a humidified chamber by placing the Petri dishescontaining the slides on test tubes that had been taped to the bottom ofa 9″×13″ Pyrex dish. These test tubes held the Petri dishes just abovethe bottom of the dish, into which about 500 mL of sterile water wasadded. The Pyrex dish was also sterilized via UV irradiation (at least 4minutes of exposure) prior to each challenge to protect againstcontamination. The dish was then covered with plastic wrap and tin foiland incubated at room temperature (20° C. to 25° C.).

Example 31 Surviving E. coli on silver Surfactant (silverbis[2-ethylhexyl]sulfosuccinate) Polymer Coated Microscope Slides

At each time point (0 hours, 12 hours, and 24 hours), 25 mL of sterilePBS was pipetted into the Petri dish completely submerging the glassslide coated with the antimicrobial polymer. A sterile plasticinoculating loop was used to release any viable bacteria remaining onthe surface of the coating, and to mix the contents of the Petri dish.Samples from this bacterial suspension were serially diluted in PBS,plated onto TSA plates, and incubated at 37° C. for at least 24 hours.Plates containing between 30 and 300 isolated colonies were counted andused to calculate concentrations (CFU/mL) of surviving bacteria. Resultsfor slides used to challenge E. coli in triplicate are illustrated inTable 14.

TABLE 14 Challenge results for AgAOT doped polyurethane coatings exposedto E. coli for 12 and 24 h on microscope slide coatings^(b). ^(a)% AgAOTsubstituted % weight for NaAOT w/w Ag CFU/mL of E. coli CFU/mL of E.coli in Coating in Coating at 12 hours^(a) at 24 hours 0 0 5.8 × 10⁵;2.9 × 10⁵; 6.8 × 10⁶; 3.4 × 10⁵; 4.1 × 10⁵ 1.2 × 10⁵ 0.00230 0.0012 6.5× 10⁴; 4.2 × 10⁴; 1.1 × 10⁵; 1.0 × 10³; 1.2 × 10⁵ 3.0 × 10³ 0.005740.0031 1.4 × 10⁵; 1.3 × 10⁵; 1.5 × 10³; 0; 3.0 × 10³ 5.8 × 10⁴ 0.01130.0061 5.0 × 10²; 1.1 × 10⁴; 0, 1.7 × 10⁴; 0 3.0 × 10⁴ 0.0226 0.012 5.5× 10⁴; 9.5 × 10³; 0, 0, 0 3.5 × 10⁴ 0.0574 0.031 0, 0, 0 0, 0, 0 0.1130.061 0, 0, 0 0, 0, 0 0.226 0.122 0, 0, 0 0, 0, 0 ^(a)The total coatingcoverage averaged 20 ± 2 mg/cm²; ^(b)the bacterial concentration of theinitial inoculum (0 h) of bacteria used to inoculate the slides wascalculated to be 4.9 £ 105 § 1.1 £ 105 CFU/ml (n = 14). CFU/ml ofsurvivors from duplicate plates

The results, shown in Table 14, after 24 h confirmed the results of thelantern slide set of challenges described in Examples 27 and 28. At andabove 0.012% weight Ag relative to overall coating weight (at and above0.0025 μmol Ag/cm²), the number of viable bacteria was reduced by3-logs. The results after 12 h exposure (Table 14) showed that completekilling of the E. coli appeared to be obtained with levels above 0.031%weight Ag relative to overall coating weight (at and above 0.00597 μmolAg/cm²). These more muted results, in comparison with 24 h exposureresults, appeared consistent with an exposure effect.

Example 32 Surviving P. Aeruginosa on silver Surfactant (AgAOT) PolymerCoated Microscope Slides

At each time point (0 hours, 12 hours, and 24 hours), 25 mL of sterilePBS was pipetted into the Petri dish completely submerging the glassslide coated with the antimicrobial polymer. A sterile plasticinoculating loop was used to release any viable bacteria remaining onthe surface of the coating, and to mix the contents of the Petri dish.Samples from this bacterial suspension were serially diluted in PBS,plated onto TSA plates, and incubated at 37° C. for at least 24 hours.Plates containing between 30 and 300 isolated colonies were counted andused to calculate concentrations (CFU/mL) of surviving bacteria. Resultsfor slides used to challenge P. aeruginosa in triplicate are illustratedin Table 15.

TABLE 15 Challenge results for AgAOT doped polyurethane coatings exposedto P. aeruginosa for 12 and 24 h on microscope slide coatings^(b). ^(a)%AgAOT substituted % weight for NaAOT w/w Ag CFU/mL of E. coli CFU/mL ofE. coli in Coating in Coating at 12 hours^(a) at 24 hours 0 0 5.5 × 10⁴;3.3 × 10⁵; 5.0 × 10⁴; 3.2 × 10⁵; 1.0 × 10⁵ 2.2 × 10⁵ 0.00230 0.0012 3.0× 10⁴; 9.5 × 10⁵; 0, 0, 0 2.6 × 10⁴ 0.00574 0.0031 1.1 × 10⁴; 1.2 × 10⁵;0; 0; 7.5 × 10² 3.3 × 10⁴ 0.0113 0.0061 4.8 × 10³; 2.5 × 10²; 0, 0, 06.3 × 10³ 0.0226 0.012 1.5 × 10³; 1.5 × 10³; 6.3 × 10³; 0; 5.0 × 10² 3.8× 10³ 0.0574 0.031 0, 0, 0 0, 0, 0 0.113 0.061 0, 0, 0 0, 0, 0 0.2260.122 0, 0, 0 0, 0, 0 ^(a)The total coating coverage averaged 20 ± 2mg/cm²; ^(b)the bacterial concentration of the initial inoculum (0 h) ofbacteria used to inoculate the slides was calculated to be 4.9 £ 105 §1.1 £ 105 CFU/ml (n = 14). CFU/ml of survivors from duplicate plates.

Similar to E. coli, the smallest substitutional level examined at 24 hexposure, 0.0012% weight Ag relative to overall coating weight (lessthan 0.0031 μmol Ag/cm²) appears effective at killing the P. aeruginosa,as do all the higher challenge levels. However, the levels of 0.0031 and0.012% (% weight Ag relative to coating weight) did not result incomplete eradication in each challenge, although the assayed levels ofCFU/ml in these cases were not statistically significantly differentthan zero. Resolution of the quantitative efficacy of this silverdelivery approach for P. aeruginosa may await more protracted testing,but the level of 0.03 1% weight Ag relative to coating weight (0.006μmol Ag/cm²) and above appear to kill all of the P. aeruginosa at both12 and 24 h exposure.

These results demonstrate that delivery of silver ion by silverbis[2-ethylhexyl]sulfosuccinate was effective as a bactericide at verylow levels of bacterial exposure. Silver bis[2-ethylhexyl]sulfosuccinatewas effective against both bacterial strains at overall levels of 0.06%(% weight silver relative to coating weight) with 12 hours of exposureand at 0.006% (% weight silver relative to coating weight) with 24 hoursof exposure.

Example 33 E. coli Challenged by silver Surfactants in Commercial AlkydPaint

A can of commercially available alkyd resin paint (Glidden Ultrahide OilAlkyd Semigloss white, GL-3517-0100 white, ICI Paints, Cleveland, Ohio,USA) is purchased at a paint store and used to prepare series of coatedmicroscope slides after adding various amounts of our silverbis(2-ethylhexyl)sulfosuccinate (AgAOT) described in Example 2, silverlinearalkanebenzene sulfonate (AgLABS) described in Example 4, andsilver dodecyl lsulfate (AgDS) described in Example 3. Xylene/methanol(5:1 by weight) is used to dissolve the silver surfactants to facilitatehomogeneously mixing with the alkyd resin composition.

TABLE 16 Challenges of E. coli by alkyd resin paint coating containingsilver anionic surfactants; 24 h contact time. Weight % Ag in CoatingAgAOT AgLABS AgDS 0 + + + 0.12 − − − 0.06 − − − 0.03 − − − 0.012 − + +0.006 P + + 0.003 + + + 0.0012 + + +

The coatings are challenged by E. coli as described in Example 12, andthe results of surviving bacteria are summarized in Table 16 for 24 hcontact time. A minus sign (−) denotes no growth, a plus sign (+)denotes confluent growth, and a capital P (P) denotes particle growth.It appears that all three silver surfactants of the present inventionare effective at preventing any growth at the 0.03 % w/w silver level.The AgAOT appears also effective at the lower level of 0.012% w/w, andpartially effective at the even lower level of 0.006% w/w.

Example 34 E. coli Challenged by silver Surfactants in CommercialAcrylic Paint

A can of commercially available solvent borne acrylic resin paint (PPGBellstar Acrylic Enamel, DAR400 White; Pittsburgh Plate and Glass,Pittsburgh, Pa.) is purchased at a paint store and used to prepareseries of coated microscope slides after adding various amounts of oursilver bis(2-ethylhexyl)sulfosuccinate (AgAOT) described in Example 32.Xylene/methanol (5:1 by weight) is used to dissolve the silversurfactants to facilitate homogeneously mixing with the acrylic resincomposition.

TABLE 17 Challenges of E. coli by commercial solvent borne acrylatepaint coating containing silver anionic surfactants; 24 h contact time.Weight % Ag in Coating AgAOT AgLABS AgDS 0 + + + 0.12 − − − 0.06 − − −0.03 P P − 0.012 + + + 0.006 + + + 0.003 + + + 0.0012 + + +

The coatings are used to challenge E. coli as described in Example 12,and the results of surviving bacteria are summarized in Table 17 after24 h contact time. A minus sign (−) denotes no growth, a plus sign (+)denotes confluent growth, and a capital P (P) denotes particle growth.It appears all three silver surfactants of the present invention areeffective at preventing any growth at the 0.06 % w/w silver level. TheAgAOT and AgLABS appear partially effective at the 0.03 % Ag w/w level,and AgDS appears completely effective at this lower level. It appearsthe most effective silver surfactant to use in a given formulationdepends on the details of the resin or binder-containing formulation andthe properties of the anionic surfactant used to form the silver anionicsurfactant of the present invention. Such optimization may be done bystraightforward experimentation by one of ordinary skill in the art.

Example 35 S. aureus Challenged by silver Surfactants in LaboratoryAcrylic Paint

A laboratory experimental solvent based acrylic paint is formulated andthen used to challenge S. aureus by adding various levels of theinvention silver anionic surfactants as described in Example 33. Abinder is made by solution polymerization of butyl acrylate (25% w/w),methylmethacrylate (73.5% w/w), and methacrylic acid (1.5% w/w) inamylacetate using thermal initiation. This solution polymer is then usedto make a 60% solids clearcoat binder in xylene/methanol (5:1 byweight). The silver anionic surfactants are dissolved in this samexylene/methanol mixture prior to addition to the clearcoat.

TABLE 18 Challenges of E. coli by laboratory solvent borne acrylateclearcoat containing silver anionic surfactants; 24 h contact time.Weight % Ag in Coating AgAOT AgLABS AgDS 0 + + + 0.12 − − − 0.06 − − −0.03 − − − 0.012 − P − 0.006 − P − 0.003 − − − 0.0012 P − −

The coatings are used to challenge S. aureus as described in Example 12,and the results of surviving bacteria are summarized in Table 18 after24 h contact time. It appears all three silver surfactants of thepresent invention are very effective at preventing any growth down tothe 0.0012% w/w silver level, with some exceptions for partial growthobserved at this lowest level for AgAOT and some partial growth at 0.012and 0.006% Ag w/w for AgLABS.

Example 36 E. coli Challenged by silver Surfactants in LaboratoryAcrylic Paint

The same laboratory experimental solvent based acrylic paint describedin Example 34 is used to challenge E. coli by adding various levels ofthe invention silver anionic surfactants as described in Example 34.

TABLE 19 Challenges of E. coli by laboratory solvent borne acrylatepaint coating containing silver anionic surfactants; 24 h contact time.Weight % Ag in Coating AgAOT AgLABS AgDS 0 + + + 0.12 − − − 0.06 − − −0.03 + P − 0.012 + + P 0.006 + + + 0.003 + + + 0.0012 + + +

The challenges to E. coli are done as described in Example 12, and theresults of surviving bacteria are summarized in Table 19 after 24 hcontact time. It appears all three silver surfactants of the presentinvention are very effective at preventing any growth down to the 0.06%w/w silver level. AgLABS has some inhibitory effects at the 0.03% level,since only partial growth is observed, and AgDS completely inhibitsgrowth at this 0.03% level, and partially inhibits at the lower 0.012%level.

Example 37 E. coli Challenged by silver Surfactants in Commercial 2KEpoxy Paint

A commercially available 2K epoxy resin paint is obtained from a localpaint store (Sherwin Williams High Solids Epoxy Tile Clad HS; Part AB62WC11; Part B B60VZ75, The Sherwin Williams Company) and the samesilver anionic surfactants described in Example 35 are used to preparetest coatings on microscope slides containing various levels of silverion. These surfactants are dissolved in xylene/methanol (5:1 by weight)to facilitate their quantitative addition to the coating mixture.

TABLE 20 Challenges of E. coli by commercial s2K epoxy paint coatingcontaining silver anionic surfactants; 24 h contact time. Weight % Ag inCoating AgAOT AgLABS AgDS 0 + + + 0.12 − − − 0.06 − − − 0.03 + − −0.012 + + P 0.006 + + + 0.003 + + + 0.0012 + + +

The coatings are used to challenge E. coli as described in Example 12,and the results of surviving bacteria are summarized in Table 20 after24 h contact time. It appears that the AgAOT is effective at completeinhibition of growth at the 0.06% w/w silver level. The AgLABS and AgDSprovide complete inhibition at the lower 0.03% w/w silver level, and theAgDS is partially effective at the lower 0.012% w/w silver level.

Example 38 Polyurethane Composition with silverbis(2-ethylhexyl)sulfosuccinate

Compositions of polyurethane suitable for forming Foley catheters basedon tolyldiisocyange (TDI) and dihydroxy polyproyleneglycol (PPG) areprepared by heating the TDI and PPG in the absence of air at 90° C.overnight in the present of dibutyltin diacetate as catalyst and dopedwith 0-3% by weight silver ion by incorporation of AgAOT into thecomposition during late stages of the synthesis. PU formulated at aTDI:PPG mole ratio of 1.5:1 are post treated with water. The AgAOT isdissolved in xylene when incorporated into the polymer reaction mixture.

Example 39 Silver Surfactant Containing PU Composition Used to ChallengeE. coli and S. aureus

Compositions of polyurethane (PU) containing 0, 0.01, 0.03, 0.1, 0.3,1.0, and 3.0% w/w silver ion from AgAOT are cut into 1″×3″ slabs andused to challenge both E. coli and S. aureus as described in Example 12.The duplicate challenge results of surviving bacteria are summarized inTable 21. The AgAOT incorporated into the PU appears equally effectiveagainst both bacteria and provides complete inhibition at 0.30% w/wsilver.

TABLE 21 Challenges of E. coli and S. aureus by PU containing AgAOT; 24h contact time. Weight % Ag in PU composition E. coli S. aureus 0 + +0.01 + + 0.03 + + 0.10 + + 0.30 − − 1.0 − − 3.0 − −

Example 40 Exposure of Bacteria to sodium Surfactant (sodiumbis[2-ethylhexyl]sulfosuccinate) on Agar Plates

Sodium bis[2-ethylhexyl]sulfosuccinate was obtained from FisherScientific (Hanover, Ill., U.S.A.). Plates containing 0.5% sodiumbis[2-ethylhexyl]sulfosuccinate were prepared by adding an equal volumeof filter sterilized sodium bis[2-ethylhexyl]sulfosuccinate to sterile,molten Trypticase Soy Agar (TSA) that was mixed and autoclaved at a 2×concentration. This solution was then aliquoted into sterile petridishes, approximately 25 mL per plate, and allowed to solidify. Theorganisms were grown in 5 mL of Trypticase Soy Broth. The sterile brothwas inoculated with a single isolated colony from a stock culture andallowed to incubate in a shaking water bath at 37° C. for 8-10 hours.The estimated concentration was about 109 CFU/mL. Some plates weretested with a 1:100 dilution of these cultures to arrive at anapproximate concentration of 107 CFU/mL. Then, 10 μL of each overnightculture was pipetted directly onto the surface of the surfactant/TSAplates, which were then allowed to incubate at 37° C. for 24 hours.Results were based on the presence or absence of bacterial growth on thesurface of the agar.

Growth of the bacteria was inhibited on plates containing sodiumbis[2-ethylhexyl]sulfosuccinate, but growth was sustained on plates freeof sodium bis[2-ethylhexyl]sulfosuccinate. This inhibition appeared tobe specific for Gram-positive bacteria, and it was not observed forGram-negative bacteria. All Gram-negative bacteria tested, except forSerratia marcescens, was resistant to the presence of sodiumbis[2-ethylhexyl]sulfosuccinate. All Gram-positive bacteria tested, aswell as the yeast Candida tropicalis, was killed on agar platescontaining sodium bis[2-ethylhexyl]sulfosuccinate. Results are shown inTables 22 and 23.

TABLE 22 Survival of Gram-positive bacteria (and the yeast Candidatropicalis) exposed to sodium surfactant (sodiumbis[2-ethylhexyl]sulfosuccinate) on agar plates. *No Growth(−)/FullGrowth(+) TSA with 0.5% TSA without sodium bis[2- sodium bis[2-Gram-positive ethylhexyl]sulfosuccinate ethylhexyl]sulfosuccinateBacteria Name 1* 2* 1* 2* Staphylococcus − − − − + + + + aureus Bacillussubtilis − − − − + + + + Streptococcus − − − − − − + + viridansStaphylococcus − − − − + + + + aureus Streptococcus bovis − − −− + + + + Enterococcus fecalis − − − − + + + + Staphylococcus − − −− + + + + epidermidis Streptococcus − − − − − − − − pneumoniaeStreptococcus − − − − + + + + pyogenes Streptococcus − − − − + + + +agalactiae Candida tropicalis − − − − + + + + Bacillus subtilis − − −− + + + + Micrococcus luteus − − − − + + + +

TABLE 23 Survival of Gram-positive bacteria exposed to various sodiumsurfactants, with structures shown in Table 24, on agar plates. *NoGrowth(−)/Full Growth(+) Compound # Bacterial S₁ S₂ S₃ S₄ S₅ S₆ NameStrain 1 2 1 2 1 2 1 2 3 1 2 3 1 2 3 1 2 3 Staphylococcus ATCC6538 − − −− − − − − − + + + + + + + + + aureus Bacillus subtilis ATCC33 − − − − −− − − − − P + − − P + + − Streptococcus EM8 − − − − − − +P + + + + + + + + + + viridans Staphylococcus EM24/ATCC25923 − − − − − −− − − + + + + + + + + + aureus Streptococcus EM12 − − − − − − − − − − −− − − − − − − bovis Enterococcus EM19 − − − − − − − PP + + + + + + + + + fecalis Enterococcus EM19 − − − − −− + + + + + + + + + + + + fecalis Staphylococcus EM3/ATCC12228 − − − − −− − P − + + + + + + + + + epidermidis Streptococcus EM14 − − − − − − − −− − − − − − − − − − pyogenes Streptococcus EM13 − − − − − − − − − − P +− − + − + − agalactiae Bacillus subtilis EM27 − − − − − − − − P P P + −P + − + − Micrococcus EM17 − − − − − − P P − − − + + − − + + + luteus

TABLE 24 Chemical structures of compounds to which Gram-positivebacteria were exposed, as in Table 23.

Not all of the Gram-positive bacteria were eradicated equally by thesurfactants evaluated as antimicrobial agents. Efficacy of thesurfactants as antimicrobial agents varied with effectivehydrophobicity, or C×log P, of the surfactant, where P is the calculatedpartition coefficient of the surfactant between 1-octanol and water, andwhere C is the concentration of the surfactant. Compounds 2 and 20 wereeffective antimicrobial agents against all Gram-positive bacteriatested. The relative ranking of hydrophobicity, and activity asantibacterial agents for Gram-positive bacteria, was the following forthe compounds: 5˜6<2˜20<4<3.

Example 41 Exposure of Bacteria to sodium Surfactant Polymer CoatedSlides

Microscope slides with polymeric coatings containing the sodiumsurfactant of sodium bis[2-ethylhexyl]sulfosuccinate were preparedaccording to the following, except all slides contained 30% (by weight)sodium bis[2-ethylhexyl]sulfosuccinate and no silverbis[2-ethylhexyl]sulfosuccinate: Glass lantern slides (2×3 inches,Fisher Scientific, Hanover, Ill., U.S.A.) and microscope slides (1×3inches, Fisher Scientific, Hanover, Ill., U.S.A.) were sterilized to ridthem of any contaminating microorganisms by exposing the slides to agermicidal UV lamp in a Nuaire Model NU-455-600 Class II Type A/B3laminar flow hood (2 minute-exposure on each side at room temperature).The glass slides were then washed with acetone and water.

Polyurethane coatings were formed on the glass slides in situ usingmicroemulsion polymerization as described in Texter et al. supra. Themicroemulsions were formulated with equivalent weights of the immisciblestep-polymerization monomers propylene glycol (PG) and isophoronediisocyanate (IPDI), along with sodium bis[2-ethylhexyl]sulfosuccinate(as described in Example 1) at 30% (% weight relative to weight of allthree components). Dibutyltin dilaurate (DBTD) was used as a catalyst todrive the step polymerization, and it was added to 0.1% of the totalcomposition weight. Coatings of approximately 150 μm thickness and 5 cmwidth were made on glass slides using a drawdown bar. The coatings werestored for at least ten days a room temperature before being subjectedto challenge testing. These sodium bis[2-ethylhexyl]sulfosuccinatecoatings were used as control coatings in the subsequent challenges.Silver bis[2-ethylhexyl]sulfosuccinate (prepared as described inExample 1) was substituted for sodium bis[2-ethylhexyl]sulfosuccinate ona weight basis (of the total composition) from 10% to 0.01% by weight,during the microemulsion formation stage prior to coating, to producetest coatings.

Sodium bis[2-ethylhexyl]sulfosuccinate killed all of the B. subtilis andS. aureus after 12 and 24 hours of exposure (Table 25).

TABLE 25 Survival of bacteria exposed to sodium surfactant (sodiumbis[2- ethylhexyl]sulfosuccinate) polyurethane coated microscope slides.Two data sets are shown. Exposure CFU/mL CFU/mL (hours) B. subtilis S.aureus 0 3.6 × 10⁴ 2.0 × 10⁵ 1.6 × 10⁴ 5.1 × 10⁵ 12 0 0 0 0 24 0 0 0 0

While the compositions and methods of this invention have been describedin terms of preferred embodiments, it will be apparent to those of skillin the art that variations may be applied to the compositions andmethods and in the steps or in the sequence of steps of the methoddescribed herein without departing from the concept, spirit and scope ofthe invention. More specifically, it will be apparent that certainagents which are both chemically and physiologically related may besubstituted for the agents described herein while the same or similarresults would be achieved. All such similar substitutes andmodifications apparent to those skilled in the art are deemed to bewithin the spirit, scope and concept of the invention.

1. A method for making a bactericidal coating comprising: (a) dissolvinga silver ion monodentate ligand complex in a suitable solvent to producea silver ion containing solution, the ligand being selected from thegroup consisting of anionic surfactants and anionic polyelectrolytes;(b) applying said solution to a suitable substrate; and (c) drying toyield a coating.
 2. The method of claim 1, wherein said substrate isselected from the group consisting of mammalian tissue, wound dressingmaterials, fabrics, synthetic absorbent materials, clothing articles,ceilings, walls, wall fixtures, ceiling fixtures, window treatments,shelving, fixed and moveable furniture, banisters, appliances,computers, machines, and filing cabinets.
 3. The method of claim 1,wherein said applying is done by spraying, pouring, metering, coating,pumping, filling, wiping, and flowing.
 4. The method of claim 1, whereinsaid anionic surfactant comprises an anionic functional group selectedfrom the group consisting of carboxylates, sulfates, sulfonates, andphosphates.
 5. The method of claim 1, wherein said anionic surfactant isselected from the group consisting of soaps, ether carboxylic acids,alkylarylsulfonates, alkane sulfonates, olefin sulfonates,sulfosuccinate diesters, sulfosuccinate monoesters, sulfosuccinimates,sulfo gluconate diesters, alcohol sulfates, alcohol ether sulfates,sulfated glycerides, sulfated alkanol amides, isethionates, taurates,sarcosinates, N-acyl amino acids, α-sulfo fatty acid methyl esters,phosphoric acid monoesters, phosphoric acid diesters, and anionic Geminisurfactants.
 6. The method of claim 1, wherein said polyelectrolytes arethe group of polyelectrolytes comprising anionic functional groupsselected from the group consisting of carboxylates, sulfates,sulfonates, and phosphates.
 7. A method for making a bactericidalcoating comprising: (a) dissolving a silver ion mondentate ligandcomplex in a suitable solvent to produce a silver ion containingsolution, the ligand being selected from the group consisting of anionicsurfactants and anionic polyelectrolytes; (b) mixing said solution witha fluid coating composition comprising a binder selected from the groupconsisting of polymers and polymer precursor monomers to produce asilver containing composition; (c) applying said silver containingcomposition to a suitable substrate to produce a silver containingcoating; and (d) curing said silver containing coating.
 8. The method ofclaim 7, wherein said anionic surfactant comprises an anionic functionalgroup selected from the group consisting of carboxylates, sulfates,sulfonates, and phosphates.
 9. The method of claim 7, wherein saidanionic surfactant is selected from the group consisting of soaps, ethercarboxylic acids, alkylarylsulfonates, alkane sulfonates, olefinsulfonates, sulfosuccinate diesters, sulfosuccinate monoesters,sulfosuccinimates, sulfo gluconate diesters, alcohol sulfates, alcoholether sulfates, sulfated glycerides, sulfated alkanol amides,isethionates, taurates, sarcosinates, N-acyl amino acids, α-sulfo fattyacid methyl esters, phosphoric acid monoesters, phosphoric aciddiesters, and anionic Gemini surfactants.
 10. The method of claim 7,wherein said polyelectrolytes are the group of polyelectrolytescomprising anionic functional groups selected from the group consistingof carboxylates, sulfates, sulfonates, and phosphates.
 11. The method ofclaim 7, wherein said fluid coating composition is a paint comprising abinder.
 12. The method of claim 7, wherein said fluid coatingcomposition comprises prepolymers and suitable cross-linking agents. 13.A bactericidal composition comprising: (a) a silver ion mondentateligand complex, the ligand being selected from the group consisting ofanionic surfactants and anionic polyelectrolytes; and (b) a polymericbinder.
 14. The composition of claim 13, wherein said anionic surfactantcomprises an anionic functional group selected from the group consistingof carboxylates, sulfates, sulfonates, and phosphates.
 15. Thecomposition of claim 13, wherein said anionic surfactant is selectedfrom the group consisting of soaps, ether carboxylic acids,alkylarylsulfonates, alkane sulfonates, olefin sulfonates,sulfosuccinate diesters, sulfosuccinate monoesters, sulfosuccinimates,sulfo gluconate diesters, alcohol sulfates, alcohol ether sulfates,sulfated glycerides, sulfated alkanol amides, isethionates, taurates,sarcosinates, N-acyl amino acids, α-sulfo fatty acid methyl esters,phosphoric acid monoesters, phosphoric acid diesters, and anionic Geminisurfactants.
 16. The composition of claim 13, wherein saidpolyelectrolytes are the group of polyelectrolytes comprising anionicfunctional groups selected from the group consisting of carboxylates,sulfates, sulfonates, and phosphates.
 17. A method for making abactericidal composition comprising: (a) dissolving a silver ionmondentate ligand complex in a suitable solvent to produce a silver ioncontaining solution, ligand being selected from the group consisting ofanionic surfactants and anionic polyelectrolytes; (b) mixing saidsolution with a composition comprising binder selected from the groupconsisting of polymers and polymer precursor monomers to produce asilver containing composition; and (c) curing said silver containingcomposition.
 18. The method of claim 17, wherein said anionic surfactantcomprises an anionic functional group selected from the group consistingof carboxylates, sulfates, sulfonates, and phosphates.
 19. The method ofclaim 17, wherein said anionic surfactant is selected from the groupconsisting of soaps, ether carboxylic acids, alkylarylsulfonates, alkanesulfonates, olefin sulfonates, sulfosuccinate diesters, sulfosuccinatemonoesters, sulfosuccinimates, sulfo gluconate diesters, alcoholsulfates, alcohol ether sulfates, sulfated glycerides, sulfated alkanolamides, isethionates, taurates, sarcosinates, N-acyl amino acids,α-sulfo fatty acid methyl esters, phosphoric acid monoesters, phosphoricacid diesters, and anionic Gemini surfactants.
 20. The method of claim17, wherein said polyelectrolytes are the group of polyelectrolytescomprising anionic functional groups selected from the group consistingof carboxylates, sulfates, sulfonates, and phosphates.